subject:(Polymerase chain reaction)

University of Waikato

1. Anderson, Michael Andrew. Detection of Toxoplasma Gondii in Fresh New Zealand Farmed Meat by the Polymerase Chain Reaction .

Degree: 2015, University of Waikato

URL: http://hdl.handle.net/10289/10118

► Toxoplasma gondii is an obligate intracellular parasite that is estimated to infect one third of the world’s human population. Upon infection this parasite causes the… (more)

▼ Toxoplasma gondii is an obligate intracellular parasite that is estimated to infect one third of the world’s human population. Upon infection this parasite causes the disease toxoplasmosis which in most healthy humans is chronic and often asymptomatic. If infection occurs in a human host without a competent immune system such as an unborn baby or infant or someone suffering from AIDS, toxoplasmosis can lead to permanent disability or death. A growing amount of evidence indicates a statistical relationship between chronic infection in healthy adults and neurological disorders leading to behavioural and psychological problems. Toxoplasma gondii can be spread through contact with cat faeces. This was thought to be the main mode of human infection. Subsequent testing with murine and Felidae models has shown that the bradyzoite of Toxoplasma gondii within intracellular cysts in muscle and neural tissue are far more infectious than the oocyst morphology shed in cat faeces. This indicates that consumption of infected meat products may also play a major contributing role in the world’s high rate of toxoplasmosis infection. The chief aim of this study was to develop a sensitive PCR assay to detect the presence of Toxoplasma gondii in fresh raw meat products. Beef, chicken, lamb and pork mince were chosen as convenient forms of fresh raw meat as each pack available for purchase contains the meat from several animal’s tissues (increasing the likelihood of contamination) and it has been partially homogenised allowing for easier sampling and DNA extraction. Secondary aims of this study were to evaluate PCR primer sets targeting polymorphic regions to gain some understanding of what strains of Toxoplasma gondii are present in our meat and the prevalence in the local cat populations. To accomplish these aims positive and negative PCR controls were also developed using a live toxoplasmosis vaccine and DNA extracted from Zebrafish. DNA extraction techniques were developed to purify DNA from both muscle tissue and feline faecal samples and primer sets were selected from published literature as well as designed using NCBI primer BLAST. Toxoplasma gondii specific DNA was detected in only one of the 25 meat samples purchased. This was achieved using a primer set targeting the P30 region of the SAG1 gene and confirmed by DNA sequencing. Using primer sets targeting polymorphic sites within the Toxoplasma gondii genome, sequencing and restriction enzyme digests it was shown that Toxoplasma gondii present in the feral Felis catus population in the raglan area is type II at the GRA6 and SAG3 loci. Amplification of polymorphic PCR targets from DNA extracted from meat samples could not be achieved. Non-specific amplification occurred with a majority of primer sets trialed with meat DNA extracts indicating improvements are needed in primer design for the purpose of PCR detection of Toxoplasma gondii in meat. Nested PCR increased specificity however resulted in contamination in several instances. Advisors/Committee Members: Cursons, Raymond T (advisor), Peters, Linda (advisor).

Subjects/Keywords: Toxoplasma; Polymerase Chain Reaction

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APA (6^th Edition):

Anderson, M. A. (2015). Detection of Toxoplasma Gondii in Fresh New Zealand Farmed Meat by the Polymerase Chain Reaction . (Masters Thesis). University of Waikato. Retrieved from http://hdl.handle.net/10289/10118

Chicago Manual of Style (16^th Edition):

Anderson, Michael Andrew. “Detection of Toxoplasma Gondii in Fresh New Zealand Farmed Meat by the Polymerase Chain Reaction .” 2015. Masters Thesis, University of Waikato. Accessed April 10, 2021. http://hdl.handle.net/10289/10118.

MLA Handbook (7^th Edition):

Anderson, Michael Andrew. “Detection of Toxoplasma Gondii in Fresh New Zealand Farmed Meat by the Polymerase Chain Reaction .” 2015. Web. 10 Apr 2021.

Vancouver:

Anderson MA. Detection of Toxoplasma Gondii in Fresh New Zealand Farmed Meat by the Polymerase Chain Reaction . [Internet] [Masters thesis]. University of Waikato; 2015. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10289/10118.

Council of Science Editors:

Anderson MA. Detection of Toxoplasma Gondii in Fresh New Zealand Farmed Meat by the Polymerase Chain Reaction . [Masters Thesis]. University of Waikato; 2015. Available from: http://hdl.handle.net/10289/10118

Indian Institute of Science

2. Mondal, Sudip. Development And Optimization Of A Microchip PCR System Using Fluorescence Detection.

Degree: PhD, Faculty of Science, 2011, Indian Institute of Science

URL: http://etd.iisc.ac.in/handle/2005/1073

► Microfabricated thermal cyclers for nucleic acid amplification by using polymerase chain reaction (PCR) have been demonstrated by several groups over the last decade, with improved… (more)

▼ Microfabricated thermal cyclers for nucleic acid amplification by using polymerase chain reaction (PCR) have been demonstrated by several groups over the last decade, with improved cycling speed and smaller volumes when compared to conventional bench-top cyclers. However, high fabrication costs coupled with difficulties in temperature sensing and control remain impediments to commercialization. In this study we have used a silicon-glass device that takes advantage of the high thermal conductivity of silicon but at the same time utilizes minimum number of fabrication steps to make it suitable for disposable applications. The thermal cycler is based on noncontact induction heating developed in this group. The microchip reaction kinetics is studied for the first time in-situ during PCR, using a real-time fluorescence block that is capable of data acquisition every 0.7 s from the microchip. The fluorescence information from SYBR green I dye is used to optimize microchip amplification reactions and confirm the product by melting curve analysis. We have also developed a novel non-contact temperature sensing technique using SYBR green fluorescence that can be used for miniaturized PCR devices. The thesis is organized into the following chapters. In chapter 1 we introduce the basic biology ideas that are required to understand DNA amplification. DNA based analysis requires amplification of low initial concentrations to above detectable limits using a technique known as polymerase chain reaction (PCR). In this process, the sample is cycled through three thermal steps for 3040 times to produce multiple copies of DNA. In microchip PCR, conventional polypropylene tubes using 2050 µL volume are replaced by miniaturized devices using ~1 µL sample volumes. The device response improves in terms of ramp rate and total analysis time due to the small volume and smart design of the materials. In this chapter we summarize some of the issues important for miniaturized PCR devices and compare them with commercial tube PCR systems. In chapter 2 we describe the induction heating technique that was developed by our group for miniaturized devices. Induction heating is a noncontact heating technique unlike resistive heating which has been commonly used for microchip PCR. Though resistive heating is very efficient in terms of heat transfer efficiency, it is not suitable for disposable devices and requires multi-step microfabrication. Other non-contact heating techniques such as hot air and IR heating require larger size arrangements that are not suitable for miniaturized devices. The heating was verified by using a thermocouple soldered at the back of the secondary plate that was also used for feedback to the comparator circuit for control. The simple on-off circuit was able to control within ±0.1 ◦C with heating and cooling ramp rates of 25 ◦C/s and 2.5 ◦C/s respectively. In this chapter, we also describe the design and fabrication of the silicon-glass microchip fabricated in our lab. We have used silicon-glass hybrid device for PCR in which… Advisors/Committee Members: Venkataraman, V (advisor).

Subjects/Keywords: Polymerase Chain Reaction (PCR); Fluorescence; Microchip; Microchip Polymerase Chain Reaction; Real-time Polymerase Chain Reaction; Induction Heater; Microchip PCR; Biophysics

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APA (6^th Edition):

Mondal, S. (2011). Development And Optimization Of A Microchip PCR System Using Fluorescence Detection. (Doctoral Dissertation). Indian Institute of Science. Retrieved from http://etd.iisc.ac.in/handle/2005/1073

Chicago Manual of Style (16^th Edition):

Mondal, Sudip. “Development And Optimization Of A Microchip PCR System Using Fluorescence Detection.” 2011. Doctoral Dissertation, Indian Institute of Science. Accessed April 10, 2021. http://etd.iisc.ac.in/handle/2005/1073.

MLA Handbook (7^th Edition):

Mondal, Sudip. “Development And Optimization Of A Microchip PCR System Using Fluorescence Detection.” 2011. Web. 10 Apr 2021.

Vancouver:

Mondal S. Development And Optimization Of A Microchip PCR System Using Fluorescence Detection. [Internet] [Doctoral dissertation]. Indian Institute of Science; 2011. [cited 2021 Apr 10]. Available from: http://etd.iisc.ac.in/handle/2005/1073.

Council of Science Editors:

Mondal S. Development And Optimization Of A Microchip PCR System Using Fluorescence Detection. [Doctoral Dissertation]. Indian Institute of Science; 2011. Available from: http://etd.iisc.ac.in/handle/2005/1073

Mississippi State University

3. Gibbs, Gordon Derek. Detection of Flavobacterium columnare in tissues and pond water using real-time polymerase chain reaction.

Degree: PhD, Veterinary Medicine, College of, 2015, Mississippi State University

URL: http://sun.library.msstate.edu/ETD-db/theses/available/etd-10302015-020003/ ;

► Flavobacterium columnare, a Gram-negative rod-shaped bacterium, is the causative agent of columnaris disease in a variety of fish hosts but is of particular significance… (more)

▼ Flavobacterium columnare, a Gram-negative rod-shaped bacterium, is the causative agent of columnaris disease in a variety of fish hosts but is of particular significance to the catfish industry located in the southeastern United States. Columnaris infections are a leading cause of mortalities in catfish ponds, occurring alone or in conjunction with other diseases. Typical diagnostic methods for columnaris infections involve the use of selective media following the observation of gross signs of disease. A real-time quantitative PCR (qPCR) assay to estimate the quantity of bacteria present in environmental and tissue samples was developed and validated. The genetic variability seen in F. columnare makes detection of isolates from different genomovars (genetic groups) essential to an assay for diagnostic application. Isolates from catfish generally fall into one of two different genomovars, one being virulent to catfish, while the other genomovar is thought to be largely opportunistic. The qPCR assay described herein was designed specifically to detect F. columnare isolates from the two major genomovars most often associated with farm-raised catfish. The assay was shown specific to F. columnare, regardless of genomovar, and demonstrated sensitivity consistent with similar qPCR assays. In addition, the assay provides quantitative information, estimating the bacterial loads in fish tissue and the environment. Two different applications of the assay are presented: (1) Estimate bacterial burden in fish tissue following immersion challenges to identify variation in transmission rates between channel and blue x channel hybrid catfish, and (2) Estimate the environmental burden of F. columnare in catfish ponds over the course of a single calendar year. This assay will provide an invaluable tool for researchers and diagnosticians in expanding our understanding of F. columnare and how it interacts with the host and environment. Advisors/Committee Members: Mark Lawrence (chair), Michael Mauel (chair), Matthew J. Griffin (committee member), Terry Greenway (committee member), Peter Allen (committee member).

Subjects/Keywords: real-time polymerase chain reaction; Flavobacterium columnare

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APA (6^th Edition):

Gibbs, G. D. (2015). Detection of Flavobacterium columnare in tissues and pond water using real-time polymerase chain reaction. (Doctoral Dissertation). Mississippi State University. Retrieved from http://sun.library.msstate.edu/ETD-db/theses/available/etd-10302015-020003/ ;

Chicago Manual of Style (16^th Edition):

Gibbs, Gordon Derek. “Detection of Flavobacterium columnare in tissues and pond water using real-time polymerase chain reaction.” 2015. Doctoral Dissertation, Mississippi State University. Accessed April 10, 2021. http://sun.library.msstate.edu/ETD-db/theses/available/etd-10302015-020003/ ;.

MLA Handbook (7^th Edition):

Gibbs, Gordon Derek. “Detection of Flavobacterium columnare in tissues and pond water using real-time polymerase chain reaction.” 2015. Web. 10 Apr 2021.

Vancouver:

Gibbs GD. Detection of Flavobacterium columnare in tissues and pond water using real-time polymerase chain reaction. [Internet] [Doctoral dissertation]. Mississippi State University; 2015. [cited 2021 Apr 10]. Available from: http://sun.library.msstate.edu/ETD-db/theses/available/etd-10302015-020003/ ;.

Council of Science Editors:

Gibbs GD. Detection of Flavobacterium columnare in tissues and pond water using real-time polymerase chain reaction. [Doctoral Dissertation]. Mississippi State University; 2015. Available from: http://sun.library.msstate.edu/ETD-db/theses/available/etd-10302015-020003/ ;

Oregon State University

4. Xu, Hong, 1968-. Development of PCR-based markers for identifying grape rootstocks.

Degree: MS, Genetics, 1995, Oregon State University

URL: http://hdl.handle.net/1957/34994

► Sequence-specific PCR markers were derived from one new and eight previously identified random amplified polymorphic DNA (RAPD) markers for the purpose of identifying grape (Vitis)… (more)

Subjects/Keywords: Polymerase chain reaction

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APA (6^th Edition):

Xu, Hong, 1. (1995). Development of PCR-based markers for identifying grape rootstocks. (Masters Thesis). Oregon State University. Retrieved from http://hdl.handle.net/1957/34994

Chicago Manual of Style (16^th Edition):

Xu, Hong, 1968-. “Development of PCR-based markers for identifying grape rootstocks.” 1995. Masters Thesis, Oregon State University. Accessed April 10, 2021. http://hdl.handle.net/1957/34994.

MLA Handbook (7^th Edition):

Xu, Hong, 1968-. “Development of PCR-based markers for identifying grape rootstocks.” 1995. Web. 10 Apr 2021.

Vancouver:

Xu, Hong 1. Development of PCR-based markers for identifying grape rootstocks. [Internet] [Masters thesis]. Oregon State University; 1995. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/1957/34994.

Council of Science Editors:

Xu, Hong 1. Development of PCR-based markers for identifying grape rootstocks. [Masters Thesis]. Oregon State University; 1995. Available from: http://hdl.handle.net/1957/34994

University of Georgia

5. Henderson, Terri Lynn. Statistical modeling and analysis of the polymerase chain reaction.

Degree: 2014, University of Georgia

URL: http://hdl.handle.net/10724/29592

► Polymerase chain reaction (PCR) is a popular method of detecting the presence of a target gene. A mathematical model is developed based upon the principals… (more)

Subjects/Keywords: Polymerase Chain Reaction; Mathematical Model; Branching Process

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APA (6^th Edition):

Henderson, T. L. (2014). Statistical modeling and analysis of the polymerase chain reaction. (Thesis). University of Georgia. Retrieved from http://hdl.handle.net/10724/29592

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Henderson, Terri Lynn. “Statistical modeling and analysis of the polymerase chain reaction.” 2014. Thesis, University of Georgia. Accessed April 10, 2021. http://hdl.handle.net/10724/29592.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Henderson, Terri Lynn. “Statistical modeling and analysis of the polymerase chain reaction.” 2014. Web. 10 Apr 2021.

Vancouver:

Henderson TL. Statistical modeling and analysis of the polymerase chain reaction. [Internet] [Thesis]. University of Georgia; 2014. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10724/29592.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Henderson TL. Statistical modeling and analysis of the polymerase chain reaction. [Thesis]. University of Georgia; 2014. Available from: http://hdl.handle.net/10724/29592

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Johannesburg

6. Foxcroft, Zyta Krystyna. Assessment of the accuracy of pre-natal rhesus D typing on amniotic fluid using the polymerase chain reaction technique.

Degree: 2014, University of Johannesburg

URL: http://hdl.handle.net/10210/10654

►

M.Tech. (Biomedical Technology)

Despite the introduction of prophylactic treatment for Rh negative females, Rhesus Haemolytic Disease of the Foetus and Newborn (HDN) remains a problem.… (more)

▼

M.Tech. (Biomedical Technology)

Despite the introduction of prophylactic treatment for Rh negative females, Rhesus Haemolytic Disease of the Foetus and Newborn (HDN) remains a problem. The serological diagnosis of this disease is mainly by maternal antibody identification and titration and the estimation of the optical density deviation (ODD) at 450 nanometers of the amniotic fluid. The correlation of these two results is not always good. The advent of molecular biology techniques such as the Polymerase Chain Reaction (PCR) and the sequencing of genes heralded the start of prenatal diagnosis of genetically inherited diseases and also enabled the prediction of the Rhesus group of the foetus. It would be advantageous to be able to predict with certainty the RhD status of a foetus suspected of having HDN without subjecting the mother and foetus to the risk of multiple invasive procedures such as Chorionic Villus Sampling (CVS) and Foetal Blood Sampling(FBS). The amniocentesis performed initially on a mother suspected of carrying an affected foetus would provide the sample necessary for the extraction of foetal DNA for prenatal Rh determination. Two PCR assays were used to determine the RhD group of the foetus: one using two primers amplifying a section ofIntron 4 and the other using four primers, two specific for Exon 7 and two specific for Exon 10 of the Rh gene. In 85.7% (18/21 cases) there was complete correlation between the molecular and the serological methods for RhD determination. One White foetus presented a unique profile, that of RhD negative in both molecular assays and RhD positive serologically. In the non-White group there were discrepancies between the two molecular methods as well as between the molecular and the serological methods used. This study shows that great care should be taken in the interpretation of RhD status prenatally using molecular biology techniques especially in the non-Caucasian population of South Africa in which there are many polymorphisrns in the Rhesus blood group system. For the moment, the results should be used in conjunction with serological results and clinical parameters for the diagnosis and treatment of Rh HDN.

Subjects/Keywords: Molecular biology; Rh factor; Polymerase chain reaction

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APA (6^th Edition):

Foxcroft, Z. K. (2014). Assessment of the accuracy of pre-natal rhesus D typing on amniotic fluid using the polymerase chain reaction technique. (Thesis). University of Johannesburg. Retrieved from http://hdl.handle.net/10210/10654

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Foxcroft, Zyta Krystyna. “Assessment of the accuracy of pre-natal rhesus D typing on amniotic fluid using the polymerase chain reaction technique.” 2014. Thesis, University of Johannesburg. Accessed April 10, 2021. http://hdl.handle.net/10210/10654.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Foxcroft, Zyta Krystyna. “Assessment of the accuracy of pre-natal rhesus D typing on amniotic fluid using the polymerase chain reaction technique.” 2014. Web. 10 Apr 2021.

Vancouver:

Foxcroft ZK. Assessment of the accuracy of pre-natal rhesus D typing on amniotic fluid using the polymerase chain reaction technique. [Internet] [Thesis]. University of Johannesburg; 2014. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10210/10654.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Foxcroft ZK. Assessment of the accuracy of pre-natal rhesus D typing on amniotic fluid using the polymerase chain reaction technique. [Thesis]. University of Johannesburg; 2014. Available from: http://hdl.handle.net/10210/10654

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Univerzitet u Beogradu

7. Strelić, Nataša J., 1964-. Značaj molekularne detekcije bakterija u proceni efikasnosti terapije Rajterovog sindroma.

Degree: Biološki fakultet, 2016, Univerzitet u Beogradu

URL: https://fedorabg.bg.ac.rs/fedora/get/o:10683/bdef:Content/get

Biologija - Molekularna biologija / Biology - Molecular biology

Nalaz bakterija u sinoviji i sinovijskoj tečnosti inflamiranog zgloba je pobudilo nadu da bi antibiotska terapija mogla biti terapija izbora kod bolesnika sa Rajterovim sindromom...

Advisors/Committee Members: Magić, Zvonko.

Subjects/Keywords: Reiter's syndrome; therapy; polymerase chain reaction

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APA (6^th Edition):

Strelić, Nataša J., 1. (2016). Značaj molekularne detekcije bakterija u proceni efikasnosti terapije Rajterovog sindroma. (Thesis). Univerzitet u Beogradu. Retrieved from https://fedorabg.bg.ac.rs/fedora/get/o:10683/bdef:Content/get

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Strelić, Nataša J., 1964-. “Značaj molekularne detekcije bakterija u proceni efikasnosti terapije Rajterovog sindroma.” 2016. Thesis, Univerzitet u Beogradu. Accessed April 10, 2021. https://fedorabg.bg.ac.rs/fedora/get/o:10683/bdef:Content/get.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Strelić, Nataša J., 1964-. “Značaj molekularne detekcije bakterija u proceni efikasnosti terapije Rajterovog sindroma.” 2016. Web. 10 Apr 2021.

Vancouver:

Strelić, Nataša J. 1. Značaj molekularne detekcije bakterija u proceni efikasnosti terapije Rajterovog sindroma. [Internet] [Thesis]. Univerzitet u Beogradu; 2016. [cited 2021 Apr 10]. Available from: https://fedorabg.bg.ac.rs/fedora/get/o:10683/bdef:Content/get.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Strelić, Nataša J. 1. Značaj molekularne detekcije bakterija u proceni efikasnosti terapije Rajterovog sindroma. [Thesis]. Univerzitet u Beogradu; 2016. Available from: https://fedorabg.bg.ac.rs/fedora/get/o:10683/bdef:Content/get

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Tartu University

8. Kõressaar, Triinu. Improvement of PCR primer design for detection of prokaryotic species .

Degree: 2012, Tartu University

URL: http://hdl.handle.net/10062/25917

► Polümeraasi ahelreaktsioon ehk PCR on molekulaarbioloogia meetod, mis võimaldab paljundada spetsiifilist DNA lõiku. Protsess toimub tsükliliselt ja kätkeb endas järgmisi etappe: kaheahelalise DNA järjestuse (sihtmärkjärjestuse)… (more)

▼ Polümeraasi ahelreaktsioon ehk PCR on molekulaarbioloogia meetod, mis võimaldab paljundada spetsiifilist DNA lõiku. Protsess toimub tsükliliselt ja kätkeb endas järgmisi etappe: kaheahelalise DNA järjestuse (sihtmärkjärjestuse) sulatamist kõrgel temperatuuril, kahe spetsiifilise DNA järjestuse (PCRi praimeri) seondumist sihtmärkjärjestusele konkreetsest PCRi katsest sõltuval madalamal temperatuuril (ehk praimerite sulamistemperatuuril Tm) ning seondunud praimerite pikendamist vastavalt spetsiifilisele DNA lõigu järjestusele kindla valgulise ensüümi abil. Paljundatud DNA-d detekteeritakse, kas spetsiifiliselt geelil pikkuse järgi või reaalajas produkti paljundamise käigus tekkiva signaali abil. PCR võimaldab sel viisil tuvastada suvalisest DNA proovist (kliiniline-, veterinaar-, toidu-, keskkonnaproov jne) vaid kindlale liigile iseäralikku DNA järjestust ning seetõttu on tehnoloogia leidnud rakendust erinevates valdkondades erinevate liikide või tüvede tuvastamiseks. Üks olulisemaid eeldusi edukaks PCRi teostamiseks on täpsete ja tundlikke praimerite disainimine (PCRi praimeridisain). PCRi praimeridisain sisaldab endas erinevaid etappe muuhulgas sihtmärkjärjestuse välja valimist (nt kindla järjestuse valimist bakterigenoomist) ning PCRi praimerijärjestuste disaini. Käesolev töö ongi keskendunud PCRi praimeridisaini erinevate etappide parendamisele: laialdaselt kasutusel oleva PCRi praimeridisaini programmi Primer3-e poolt kasutatava praimerite sulamistemperatuuri Tm arvutamise valemi täiustamisele ning sellele, kuidas prokarüootseid liigispetsiifilisi kordusjärjestusi PCRi sihtmärkjärjestustena kasutada ning, millise effekti see PCRi tulemustele annab. Viimase osana sisaldab töö prokarüootsete liigispetsiifiliste kordusjärjestuste iseloomustamist 613 erinevas prokarüootses liigis. Antud töö tulemused aitavad kaasa uute ja paremate molekulaardiagnostika testide loomisele esiteks lihtsustades nende väljatöötamist tänu antud töös leitud ja kirjeldatud PCRi sihtmärkjärjestustele, teiseks tõstes nende töökindlust, kuna liigispetsiifilistele kordusjärjestustele disainitud praimerid on kõrgendatud tundlikkusega ning, kuna parendatud praimeridisaini-programm Primer3 võimaldab disainida praimereid, mis vastavad täpsemalt etteantud molekulaardiagnostika testide tingimustele .; Polymerase chain reaction or PCR is a method in molecular biology, which enables amplification of specific DNA regions. It is a cyclic process, that comprises of heat denaturation of double-stranded DNA (target sequence), hybridization of two short oligonucleotides (called PCR primers) to the denaturated target sequence at temperature specific to the PCR reaction (termed melting temperature Tm) and extension of hybridized primers by the enzyme DNA polymerase. The amplified DNA can be identified either specifically on electrophoresis gel by its product length or by monitoring the signal generated through activation of primer attached flourecent labels during the amplification in real-time. Thus, PCR enables identification of the DNA sequence of…

Subjects/Keywords: polümeraasahelreaktsioon; molekulaardiagnostika; polymerase chain reaction; molecular diagnostics

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APA (6^th Edition):

Kõressaar, T. (2012). Improvement of PCR primer design for detection of prokaryotic species . (Thesis). Tartu University. Retrieved from http://hdl.handle.net/10062/25917

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Kõressaar, Triinu. “Improvement of PCR primer design for detection of prokaryotic species .” 2012. Thesis, Tartu University. Accessed April 10, 2021. http://hdl.handle.net/10062/25917.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Kõressaar, Triinu. “Improvement of PCR primer design for detection of prokaryotic species .” 2012. Web. 10 Apr 2021.

Vancouver:

Kõressaar T. Improvement of PCR primer design for detection of prokaryotic species . [Internet] [Thesis]. Tartu University; 2012. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10062/25917.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Kõressaar T. Improvement of PCR primer design for detection of prokaryotic species . [Thesis]. Tartu University; 2012. Available from: http://hdl.handle.net/10062/25917

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Missouri – Columbia

9. Liu, Yuejiao. High Resolution Meltcurve PCR assay for detection of Salmonella and Escherichia coli o157:h7 in foods.

Degree: 2017, University of Missouri – Columbia

URL: http://hdl.handle.net/10355/62057

► Foodborne illnesses associated with Salmonella and Escherichia coli O157:H7 have become world-wide public-health problems. Conventional methods for the identification of foodborne pathogens are tedious, expensive,… (more)

Subjects/Keywords: Salmonella – Detection; Escherichia coli; Polymerase chain reaction

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APA (6^th Edition):

Liu, Y. (2017). High Resolution Meltcurve PCR assay for detection of Salmonella and Escherichia coli o157:h7 in foods. (Thesis). University of Missouri – Columbia. Retrieved from http://hdl.handle.net/10355/62057

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Liu, Yuejiao. “High Resolution Meltcurve PCR assay for detection of Salmonella and Escherichia coli o157:h7 in foods.” 2017. Thesis, University of Missouri – Columbia. Accessed April 10, 2021. http://hdl.handle.net/10355/62057.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Liu, Yuejiao. “High Resolution Meltcurve PCR assay for detection of Salmonella and Escherichia coli o157:h7 in foods.” 2017. Web. 10 Apr 2021.

Vancouver:

Liu Y. High Resolution Meltcurve PCR assay for detection of Salmonella and Escherichia coli o157:h7 in foods. [Internet] [Thesis]. University of Missouri – Columbia; 2017. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10355/62057.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Liu Y. High Resolution Meltcurve PCR assay for detection of Salmonella and Escherichia coli o157:h7 in foods. [Thesis]. University of Missouri – Columbia; 2017. Available from: http://hdl.handle.net/10355/62057

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

NSYSU

10. Chien, Hua-yi. Use of In Situ Bioremediation to Treat Trichloroethylene-contaminated Groundwater.

Degree: PhD, Environmental Engineering, 2010, NSYSU

URL: http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0207110-212744

► Chlorinated aliphatic hydrocarbons (CAHs) include tetrachloroethene (PCE), trichloroethene (TCE), and others. The industrial solvent TCE is among the most ubiquitous chlorinated compounds found in groundwater… (more)

▼ Chlorinated aliphatic hydrocarbons (CAHs) include tetrachloroethene (PCE), trichloroethene (TCE), and others. The industrial solvent TCE is among the most ubiquitous chlorinated compounds found in groundwater pollution. TCE in environment can be removed by physical, chemical and biological procedures. Dehalorespiration is a biological pathway from which bacteria can derive energy from the reductive dechlorination of chlorinated ethenes using hydrogen or organic acids as electron donors and yielding chloride and ethene as degradation products. Dehalorespiration can be used to remediate chlorinated ethene contaminated aquifers if an appropriate aquifer ecosystem exists including populations of dechlorinating bacteria and companion organisms that contribute to the biogeochemical environment conducive to dehalorespiration activity. Enhanced in-situ aerobic or anaerobic bioremediation of chlorinated solvents is a cost-effective, expanding technology for the clean-up of chlorinated solvent-contaminated sites. The objective of this pilot-scale study was to apply an enhanced in situ bioremediation technology to remediate TCE-contaminated groundwater. Both aerobic and anaerobic remedial systems were evaluated at a TCE-spill site located in southern Taiwan. In the aerobic bioremediation zone, the effectiveness of air, nutrient, and sugarcane molasses injection to enhance the aerobic cometabolism on TCE degradation was evaluated. Results show that the decreases in TCE concentration were observed over 204 days operating period. Up to 73ï¼-99ï¼ of TCE removal efficiency was obtained in this treatment system. In the anaerobic test zone, the effectiveness of nutrient and sugarcane molasses injection to enhance the anaerobic dechlorination on TCE degradation was also evaluated. Results show that the decreases in TCE concentration were observed over a 193-day operating period. Up to 53ï¼-91ï¼ of TCE removal efficiency was obtained in this treatment system. Polymerase chain reaction was applied to analyze the gene variation in TCE-microbial degraders during the treatment process. Results from this study indicate that the aerobic TCE-degraders (typeâ methanotrophs and type â¡ methanotrophs) and the gene of degradation enzymes (toluene monooxygenase, toluene dioxygenase, particulate methane monooxygenase) were detected after the treatment process in the aerobic test zone. In the anaerobic treatment zone, Dehalococcoides (anaerobic TCE-degrader) and the gene of degradation enzyme (vcrA and tceA) were detected and a significant drop of TCE concentration was also observed. Based on 16S rDNA sequence analysis, samples of groundwater from aerobic/anaerobic bioremediation zone are close related to the genera of Dehalococcoides sp. MB, Dehalococcoides ethenogenes 195, Dehalococcoides sp. VS, Acidovorax sp., Alicycliphilus sp., Burkholderiales, Caulobacter sp., Caulobacter tuntrae, Caulobacter vibrioides, Comamonadaceae, Hydrogenophaga sp., Iron-reducing bacterium, Mitsuaria chitosanitabida, Rhodocyclacea, Pseudomonas sp., Rhodoferax… Advisors/Committee Members: Huang Wen-Yen (chair), Chen Ku-Fan (chair), Yu Kuang-Chung (chair), Lai Chin-Hsing (chair), Kao Chih-Ming (committee member), Tsai Li-Jyur (chair).

Subjects/Keywords: in situ bioremediation; polymerase chain reaction; groundwater pollution; trichloroethene

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APA (6^th Edition):

Chien, H. (2010). Use of In Situ Bioremediation to Treat Trichloroethylene-contaminated Groundwater. (Doctoral Dissertation). NSYSU. Retrieved from http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0207110-212744

Chicago Manual of Style (16^th Edition):

Chien, Hua-yi. “Use of In Situ Bioremediation to Treat Trichloroethylene-contaminated Groundwater.” 2010. Doctoral Dissertation, NSYSU. Accessed April 10, 2021. http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0207110-212744.

MLA Handbook (7^th Edition):

Chien, Hua-yi. “Use of In Situ Bioremediation to Treat Trichloroethylene-contaminated Groundwater.” 2010. Web. 10 Apr 2021.

Vancouver:

Chien H. Use of In Situ Bioremediation to Treat Trichloroethylene-contaminated Groundwater. [Internet] [Doctoral dissertation]. NSYSU; 2010. [cited 2021 Apr 10]. Available from: http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0207110-212744.

Council of Science Editors:

Chien H. Use of In Situ Bioremediation to Treat Trichloroethylene-contaminated Groundwater. [Doctoral Dissertation]. NSYSU; 2010. Available from: http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0207110-212744

University of Pretoria

11. Bagla, Victor Patrick. The demonstration of lumpy skin disease virus in semen of experimentally infected bulls using different diagnostic techniques .

Degree: 2008, University of Pretoria

URL: http://upetd.up.ac.za/thesis/available/etd-05272008-120446/

► Lumpy skin disease virus (LSDV), a poxvirus that belongs to the genus Capripoxvirus is an important pathogen that can be shed in the semen of… (more)

▼ Lumpy skin disease virus (LSDV), a poxvirus that belongs to the genus Capripoxvirus is an important pathogen that can be shed in the semen of infected bulls. The screening of semen for infectious virus prior to artificial insemination requires a sensitive diagnostic method. The isolation of the virus on cell cultures and/or the use of polymerase chain reaction (PCR) are sensitive diagnostic tests which can be used to screen semen for LSD viral DNA prior to artificial insemination. Although cell culture is a sensitive method and detects infectious virus, its use has major limitations due to the toxic effect of semen on the cells. This study was therefore aimed at finding a method that decreases the toxic effect of semen on cell culture and enhances LSDV isolation. Secondly, the efficiency of this method in enhancing the isolation of LSDV in field samples was tested. In order to eliminate the toxic effect of semen on cell culture, a pilot study was conducted in which semen samples from LSDV sero-negative bulls were collected and infected with a field isolate of LSDV, strain 248/93 with a titre of 6.5 log TCID50. The semen samples were subjected to one of four different methods, viz centrifugation, serial dilution, filtration and chemical treatment with kaolin. The centrifugation, serial dilution, and filtration methods were supplemented with additional amounts of gentamycin. The toxic effects of semen on cell culture were completely eliminated when supernatants of semen samples, centrifuged at 2000 rpm for 1, 3 and 5 mins and serial diluted was used to inoculate confluent monolayers of bovine dermis cells. Semen diluted in MEM with or without additional antibiotics was the most sensitive method of demonstrating virus at higher dilutions, followed by pellets of samples centrifuged for 1 and 3 minutes. The toxicity recorded when the pellet fraction of semen samples were centrifuged for 5 mins at 2000 rpm was comparable to results obtained from serially diluted samples supplemented with gentamycin. The use of filtration and kaolin treatment of semen samples could not remove the toxic effect of semen on cells. To evaluate the presence of LSDV in semen of experimentally infected bulls, six seronegative post-pubertal bulls housed in an insect proof facility were infected with LSDV via the intravenous route. The experimentally infected bulls were monitored for clinical sign of the disease. Two bulls showed severe, two a mild and two an inapparent infection. Blood samples were collected for virus isolation and semen samples for virus isolation and PCR. Vesicular fluid and preputial washes were also investigated for the presence of LSD viral nucleic acid using PCR. The infectious titre of the virus shed in semen of these bulls was also calculated. The incubation period in infected bulls varied from 7 to 14 days. The length of viraemia varied between groups and did not correlate with the severity of clinical disease. The virus was isolated from blood samples of bulls in the severely infected group on several occasions. Bulls in… Advisors/Committee Members: Prof E H Venter (advisor).

Subjects/Keywords: Lumpy skin disease virus; Viral nucleic acid; Polymerase chain reaction; UCTD

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APA (6^th Edition):

Bagla, V. P. (2008). The demonstration of lumpy skin disease virus in semen of experimentally infected bulls using different diagnostic techniques . (Masters Thesis). University of Pretoria. Retrieved from http://upetd.up.ac.za/thesis/available/etd-05272008-120446/

Chicago Manual of Style (16^th Edition):

Bagla, Victor Patrick. “The demonstration of lumpy skin disease virus in semen of experimentally infected bulls using different diagnostic techniques .” 2008. Masters Thesis, University of Pretoria. Accessed April 10, 2021. http://upetd.up.ac.za/thesis/available/etd-05272008-120446/.

MLA Handbook (7^th Edition):

Bagla, Victor Patrick. “The demonstration of lumpy skin disease virus in semen of experimentally infected bulls using different diagnostic techniques .” 2008. Web. 10 Apr 2021.

Vancouver:

Bagla VP. The demonstration of lumpy skin disease virus in semen of experimentally infected bulls using different diagnostic techniques . [Internet] [Masters thesis]. University of Pretoria; 2008. [cited 2021 Apr 10]. Available from: http://upetd.up.ac.za/thesis/available/etd-05272008-120446/.

Council of Science Editors:

Bagla VP. The demonstration of lumpy skin disease virus in semen of experimentally infected bulls using different diagnostic techniques . [Masters Thesis]. University of Pretoria; 2008. Available from: http://upetd.up.ac.za/thesis/available/etd-05272008-120446/

University of Utah

12. Ragsdale, Victoria Elizabeth. Three-dimensional heat transfer analysis of a disposable, continuous-flow polymerase chain reaction device.

Degree: MS, Mechanical Engineering, 2012, University of Utah

URL: http://content.lib.utah.edu/cdm/singleitem/collection/etd3/id/640/rec/2741

► Polymerase Chain Reaction (PCR) is a process which amplifies a specific segment of DNA through a thermal cycling protocol. PCR is largely used in diagnostics,but… (more)

▼ Polymerase Chain Reaction (PCR) is a process which amplifies a specific segment of DNA through a thermal cycling protocol. PCR is largely used in diagnostics,but also has applications in the defense industry and many others. The PCR industry is shifting its focus towards micro-scale devices, and away from macro-scale systems due to i) the micro-scale sample size requiring less blood from patients, ii) total reaction times on the order of minutes opposed to hours and iii) the cost advantages as many microfluidic devices are manufactured from inexpensive polymers. In general, polymers have many advantages including minimal microfabrication processes during manufacturing, wide availability and low cost.This work presents the manufacturing process as well as simulations and testing results of a disposable polycarbonate (PC) device capable of achieving PCR in less than 7 minutes by thermally cycling the sample through an established temperature gradient in aserpentine channel. This device also features a unique laser manufacturing process eliminating the need for a microfabrication facility. Two-dimensional (2D) and three-dimensional (3D) simulations were created to analyze the heat transfer and fluid dynamics throughout the device. Simulations were created both in the flow plane, to analyze the reaction temperatures occurring in each bend of the serpentine channel, and the out of flow plane, to observe the amount of heat leaving the system. Although the 2D simulations did not produce the same values as the 3D models, the same trends wereexhibited so qualitative observations could still be made. Experiments were performed to validate the simulations by comparing the measured device surface temperature from an IR camera to those values from the 3D simulations. It was found that the device producesfluid temperature results within the reaction limits for several flow rates with specific heater temperature settings. An end point PCR melting analysis was also completed on reactants to determine if PCR was achieved. These results were unsuccessful, due tohighly inconsistent flow rates observed during the experiments. Although PCR was not achieved, with slight changes to increase flow rate accuracy, PCR is expected to be successful since the IR images suggest that the fluid is reaching the reaction temperatures.

Subjects/Keywords: Heat transfer; PCR; Continuous-flow; Polymerase chain reaction device

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APA (6^th Edition):

Ragsdale, V. E. (2012). Three-dimensional heat transfer analysis of a disposable, continuous-flow polymerase chain reaction device. (Masters Thesis). University of Utah. Retrieved from http://content.lib.utah.edu/cdm/singleitem/collection/etd3/id/640/rec/2741

Chicago Manual of Style (16^th Edition):

Ragsdale, Victoria Elizabeth. “Three-dimensional heat transfer analysis of a disposable, continuous-flow polymerase chain reaction device.” 2012. Masters Thesis, University of Utah. Accessed April 10, 2021. http://content.lib.utah.edu/cdm/singleitem/collection/etd3/id/640/rec/2741.

MLA Handbook (7^th Edition):

Ragsdale, Victoria Elizabeth. “Three-dimensional heat transfer analysis of a disposable, continuous-flow polymerase chain reaction device.” 2012. Web. 10 Apr 2021.

Vancouver:

Ragsdale VE. Three-dimensional heat transfer analysis of a disposable, continuous-flow polymerase chain reaction device. [Internet] [Masters thesis]. University of Utah; 2012. [cited 2021 Apr 10]. Available from: http://content.lib.utah.edu/cdm/singleitem/collection/etd3/id/640/rec/2741.

Council of Science Editors:

Ragsdale VE. Three-dimensional heat transfer analysis of a disposable, continuous-flow polymerase chain reaction device. [Masters Thesis]. University of Utah; 2012. Available from: http://content.lib.utah.edu/cdm/singleitem/collection/etd3/id/640/rec/2741

University of Utah

13. Hsieh, Fen-Fen. Development of a sensitive polymerase chain reaction assay for group A streptococci;.

Degree: MS;, Pathology;, 1995, University of Utah

URL: http://content.lib.utah.edu/cdm/singleitem/collection/etd1/id/1062/rec/308

► S. pyogenes causes many important human diseases. Rheumatic fever is a systemic inflammatory and nonsuppurative sequla of acute streptococcal pharyngitis that occurs in a small… (more)

Subjects/Keywords: Polymerase Chain Reaction; DNA

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APA (6^th Edition):

Hsieh, F. (1995). Development of a sensitive polymerase chain reaction assay for group A streptococci;. (Masters Thesis). University of Utah. Retrieved from http://content.lib.utah.edu/cdm/singleitem/collection/etd1/id/1062/rec/308

Chicago Manual of Style (16^th Edition):

Hsieh, Fen-Fen. “Development of a sensitive polymerase chain reaction assay for group A streptococci;.” 1995. Masters Thesis, University of Utah. Accessed April 10, 2021. http://content.lib.utah.edu/cdm/singleitem/collection/etd1/id/1062/rec/308.

MLA Handbook (7^th Edition):

Hsieh, Fen-Fen. “Development of a sensitive polymerase chain reaction assay for group A streptococci;.” 1995. Web. 10 Apr 2021.

Vancouver:

Hsieh F. Development of a sensitive polymerase chain reaction assay for group A streptococci;. [Internet] [Masters thesis]. University of Utah; 1995. [cited 2021 Apr 10]. Available from: http://content.lib.utah.edu/cdm/singleitem/collection/etd1/id/1062/rec/308.

Council of Science Editors:

Hsieh F. Development of a sensitive polymerase chain reaction assay for group A streptococci;. [Masters Thesis]. University of Utah; 1995. Available from: http://content.lib.utah.edu/cdm/singleitem/collection/etd1/id/1062/rec/308

14. Kolari, Kai. Fabrication of Silicon and Glass Devices for Microfluidic Bioanalytical Applications.

Degree: 2008, VTT Technical Research Centre of Finland

URL: http://lib.tkk.fi/Diss/2008/isbn9789513870720/

►

This thesis introduces important improvements in fabrication of microfluidic devices on silicon and glass. With the main aim in surface and volume manipulation of aqueous… (more)

▼

This thesis introduces important improvements in fabrication of microfluidic devices on silicon and glass. With the main aim in surface and volume manipulation of aqueous solutions for subsequent biochemical analysis, the backbone of the work has been the development of plasma etching processes for silicon and glass. As the silicon microfabrication technologies are combined with deep anisotropic etching of glass, the processability of microfluidic applications with surface and volume manipulation of fluid is diversified. Several mask materials have been studied with respect to deep plasma etching of glass. As the demand for depth of microfluidic devices extends past 150 µm, the number of usable masking schemes becomes limited. To reach an etch depth beyond 350 µm with aspect ratio of over 3:1 including the mask, silicon shadow mask was used. The results of process development on Al2O3, AlN and TiO2 masks show that a very high etching selectivity on glass can be achieved with these mask materials. The described masking technologies enable e.g. high density of through-a-wafer holes or nearly vertical structuring of glass with great depth. Also, a silicon shadow mask was used for local tuning of hydrophobicity of C4F8 polymer on silicon and glass surfaces by pattering the polymer with O2 plasma through the shadow mask. For both purposes, one silicon shadow mask wafer can be re-used to enable lower processing costs. Thermal manipulation of fluid allows polymerase chain reaction on silicon and glass microchips, but also triggering of capillary action. However, the results of a novel method indicate possible lack of biocompatibility of oxidized silicon surfaces, which may limit the usable microchip surface materials. Microfluidic components with hydrophilic patterning for controlled capillary action can be combined with microphotonics through excitation of fluorescence with evanescent field, which has been characterized with a grating-coupled laser beam.

Työssä käsitellään mikrofluidististen rakenteiden valmistamiseen liittyviä parannuksia. Työn tavoitteena on ollut nesteiden kaksi- ja kolmiulotteinen manipulointi, jonka ytimessä on plasmasyövytysprosessien kehitys piille ja lasille. Piin mikrovalmistusteknologioiden yhdistäminen lasin suurien syvyyksien anisotrooppiseen plasmasyövytykseen luo edellytyksiä uudentyyppisten mikrofluidististen sovellusten valmistamiselle. Lasin plasmasyövytykseen on sovellettu useita eri maskimateriaaleja. Pyrittäessä yli 150 µm:n plasmasyövytyssyvyyteen lasilla maskivaihtoehtojen lukumäärä on vähäinen. Pyrittäessä yli 350 µm syvyyteen ja yli 3:1 aspektisuhteeseen työssä on käytetty piivarjomaskia. Erityisesti alumiinioksidi- ja -alumiininitridimaskeilla voidaan saavuttaa hyvin suuri syöpymisnopeussuhde lasiin. Työssä kuvattuja maskiteknologioita voidaan käyttää mm. läpivientien sekä lähes pystysuorien ja erittäin syvien lasirakenteiden syövyttämiseen. Piivarjomaskiteknologia soveltuu myös käytettäväksi paikallisten hydrofiilisten kuviointien valmistamiseen hydrofobiselle…

Advisors/Committee Members: Helsinki University of Technology, Faculty of Electronics, Communications and Automation.

Subjects/Keywords: glass; plasma etching; hydrophobic coating; shadow mask; polymerase chain reaction

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APA (6^th Edition):

Kolari, K. (2008). Fabrication of Silicon and Glass Devices for Microfluidic Bioanalytical Applications. (Thesis). VTT Technical Research Centre of Finland. Retrieved from http://lib.tkk.fi/Diss/2008/isbn9789513870720/

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Kolari, Kai. “Fabrication of Silicon and Glass Devices for Microfluidic Bioanalytical Applications.” 2008. Thesis, VTT Technical Research Centre of Finland. Accessed April 10, 2021. http://lib.tkk.fi/Diss/2008/isbn9789513870720/.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Kolari, Kai. “Fabrication of Silicon and Glass Devices for Microfluidic Bioanalytical Applications.” 2008. Web. 10 Apr 2021.

Vancouver:

Kolari K. Fabrication of Silicon and Glass Devices for Microfluidic Bioanalytical Applications. [Internet] [Thesis]. VTT Technical Research Centre of Finland; 2008. [cited 2021 Apr 10]. Available from: http://lib.tkk.fi/Diss/2008/isbn9789513870720/.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Kolari K. Fabrication of Silicon and Glass Devices for Microfluidic Bioanalytical Applications. [Thesis]. VTT Technical Research Centre of Finland; 2008. Available from: http://lib.tkk.fi/Diss/2008/isbn9789513870720/

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of South Australia

15. Ellis, Geoffrey Richard. Application of the polymerase chain reaction for the rapid diagnosis of leptospirosis.

Degree: MA, 1995, University of South Australia

URL: http://arrow.unisa.edu.au:8081/1959.8/85113 ; http://arrow.unisa.edu.au/vital/access/manager/Repository/unisa:42819

Subjects/Keywords: Leptospirosis; Polymerase chain reaction.

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APA (6^th Edition):

Ellis, G. R. (1995). Application of the polymerase chain reaction for the rapid diagnosis of leptospirosis. (Masters Thesis). University of South Australia. Retrieved from http://arrow.unisa.edu.au:8081/1959.8/85113 ; http://arrow.unisa.edu.au/vital/access/manager/Repository/unisa:42819

Chicago Manual of Style (16^th Edition):

Ellis, Geoffrey Richard. “Application of the polymerase chain reaction for the rapid diagnosis of leptospirosis.” 1995. Masters Thesis, University of South Australia. Accessed April 10, 2021. http://arrow.unisa.edu.au:8081/1959.8/85113 ; http://arrow.unisa.edu.au/vital/access/manager/Repository/unisa:42819.

MLA Handbook (7^th Edition):

Ellis, Geoffrey Richard. “Application of the polymerase chain reaction for the rapid diagnosis of leptospirosis.” 1995. Web. 10 Apr 2021.

Vancouver:

Ellis GR. Application of the polymerase chain reaction for the rapid diagnosis of leptospirosis. [Internet] [Masters thesis]. University of South Australia; 1995. [cited 2021 Apr 10]. Available from: http://arrow.unisa.edu.au:8081/1959.8/85113 ; http://arrow.unisa.edu.au/vital/access/manager/Repository/unisa:42819.

Council of Science Editors:

Ellis GR. Application of the polymerase chain reaction for the rapid diagnosis of leptospirosis. [Masters Thesis]. University of South Australia; 1995. Available from: http://arrow.unisa.edu.au:8081/1959.8/85113 ; http://arrow.unisa.edu.au/vital/access/manager/Repository/unisa:42819

University of Hong Kong

16. Wong, Ting-yan, Cybil. The use of JAK2 quantitative polymerase chain reaction for the diagnosis and monitoring of patients with chronic myeloproliferativediseases.

Degree: 2008, University of Hong Kong

URL: http://hdl.handle.net/10722/52076

Subjects/Keywords: Polymerase chain reaction.; Myeloproliferative disorders.

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APA (6^th Edition):

Wong, Ting-yan, C. (2008). The use of JAK2 quantitative polymerase chain reaction for the diagnosis and monitoring of patients with chronic myeloproliferativediseases. (Thesis). University of Hong Kong. Retrieved from http://hdl.handle.net/10722/52076

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Wong, Ting-yan, Cybil. “The use of JAK2 quantitative polymerase chain reaction for the diagnosis and monitoring of patients with chronic myeloproliferativediseases.” 2008. Thesis, University of Hong Kong. Accessed April 10, 2021. http://hdl.handle.net/10722/52076.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Wong, Ting-yan, Cybil. “The use of JAK2 quantitative polymerase chain reaction for the diagnosis and monitoring of patients with chronic myeloproliferativediseases.” 2008. Web. 10 Apr 2021.

Vancouver:

Wong, Ting-yan C. The use of JAK2 quantitative polymerase chain reaction for the diagnosis and monitoring of patients with chronic myeloproliferativediseases. [Internet] [Thesis]. University of Hong Kong; 2008. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10722/52076.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Wong, Ting-yan C. The use of JAK2 quantitative polymerase chain reaction for the diagnosis and monitoring of patients with chronic myeloproliferativediseases. [Thesis]. University of Hong Kong; 2008. Available from: http://hdl.handle.net/10722/52076

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Hong Kong

17. Tsang, Tsui-ying, Stella. Application of quantitative polymerase chain reaction in the diagnosisof thalassaemia.

Degree: 2006, University of Hong Kong

URL: http://hdl.handle.net/10722/131307

Subjects/Keywords: Thalassemia - Diagnosis.; Polymerase chain reaction.

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APA (6^th Edition):

Tsang, Tsui-ying, S. (2006). Application of quantitative polymerase chain reaction in the diagnosisof thalassaemia. (Thesis). University of Hong Kong. Retrieved from http://hdl.handle.net/10722/131307

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Tsang, Tsui-ying, Stella. “Application of quantitative polymerase chain reaction in the diagnosisof thalassaemia.” 2006. Thesis, University of Hong Kong. Accessed April 10, 2021. http://hdl.handle.net/10722/131307.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Tsang, Tsui-ying, Stella. “Application of quantitative polymerase chain reaction in the diagnosisof thalassaemia.” 2006. Web. 10 Apr 2021.

Vancouver:

Tsang, Tsui-ying S. Application of quantitative polymerase chain reaction in the diagnosisof thalassaemia. [Internet] [Thesis]. University of Hong Kong; 2006. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10722/131307.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Tsang, Tsui-ying S. Application of quantitative polymerase chain reaction in the diagnosisof thalassaemia. [Thesis]. University of Hong Kong; 2006. Available from: http://hdl.handle.net/10722/131307

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Hong Kong

18. Lam, Yiu-pong. Performance evaluation of the automated NucliSens easyMAG and Qiagen EZ1 Advanced XL nucleic acid extraction platform for detection of RNAand DNA viruses in clinical samples.

Degree: 2011, University of Hong Kong

URL: http://hdl.handle.net/10722/141956

Subjects/Keywords: Polymerase chain reaction.; Molecular diagnosis.

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APA (6^th Edition):

Lam, Y. (2011). Performance evaluation of the automated NucliSens easyMAG and Qiagen EZ1 Advanced XL nucleic acid extraction platform for detection of RNAand DNA viruses in clinical samples. (Thesis). University of Hong Kong. Retrieved from http://hdl.handle.net/10722/141956

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Lam, Yiu-pong. “Performance evaluation of the automated NucliSens easyMAG and Qiagen EZ1 Advanced XL nucleic acid extraction platform for detection of RNAand DNA viruses in clinical samples.” 2011. Thesis, University of Hong Kong. Accessed April 10, 2021. http://hdl.handle.net/10722/141956.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Lam, Yiu-pong. “Performance evaluation of the automated NucliSens easyMAG and Qiagen EZ1 Advanced XL nucleic acid extraction platform for detection of RNAand DNA viruses in clinical samples.” 2011. Web. 10 Apr 2021.

Vancouver:

Lam Y. Performance evaluation of the automated NucliSens easyMAG and Qiagen EZ1 Advanced XL nucleic acid extraction platform for detection of RNAand DNA viruses in clinical samples. [Internet] [Thesis]. University of Hong Kong; 2011. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10722/141956.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Lam Y. Performance evaluation of the automated NucliSens easyMAG and Qiagen EZ1 Advanced XL nucleic acid extraction platform for detection of RNAand DNA viruses in clinical samples. [Thesis]. University of Hong Kong; 2011. Available from: http://hdl.handle.net/10722/141956

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Pretoria

19. [No author]. Development of PCR-based methods for detection of African lyssaviruses .

Degree: 2010, University of Pretoria

URL: http://upetd.up.ac.za/thesis/available/etd-10082010-143715/

► The etiological agent of rabies encephalitis belongs to the genus Lyssavirus in the Rhabdoviridae family. Lyssaviruses are negative sense, single stranded RNA viruses and cause… (more)

▼ The etiological agent of rabies encephalitis belongs to the genus Lyssavirus in the Rhabdoviridae family. Lyssaviruses are negative sense, single stranded RNA viruses and cause an estimated 55 000 human deaths per year with 44% of these deaths occurring in Africa (WHO, 2005). With intense research effort and increased sequence information it is becoming evident that the Lyssavirus genus is much more diverse than initially thought and therefore diagnostic methods need to be modified accordingly. The African continent sustains a diverse variety of lyssaviruses, however, most countries in Africa do not have active surveillance or necessary diagnostic tools and therefore rabies-related lyssaviruses are underreported. Previous studies have indicated that real-time PCR has improved sensitivity and rapidity over conventional molecular diagnostic methods with the added advantage of allowing accurate estimations of viral load in a wide variety of samples. Several realtime PCR assays have been developed; however, none were specifically aimed at detection of lyssaviruses present on the African continent. This study was therefore aimed at evaluating certain molecular diagnostic methods for the detection of African lyssaviruses. Furthermore, the application of real-time PCR for various fields in lyssavirus research i.e. diagnostics, surveillance and pathogenicity studies were evaluated. This study revealed two different hemi-nested PCR assays capable of detecting representatives of African lyssaviruses. A real-time PCR was developed that was successful for the detection of African lyssaviruses. In addition, a quantitative assay and internal control was successfully employed for confirming ante-mortem human rabies diagnosis as well as post-mortem animal rabies diagnosis in formalin fixed brain material. As such the real-time PCR assay developed in this study could therefore be routinely used for ante-mortem diagnosis and as a confirmatory test for post-mortem diagnosis. The ability of this assay to detect and quantify all currently known African lyssaviruses not only offers improved surveillance capacity, but offers unique potential as a sensitive tool to track virus movement in pathogenicity studies. These aspects are important in our search for a better understanding of the complex epidemiological and viral characteristics of African lyssaviruses. Copyright Advisors/Committee Members: Weyer, Jacqueline (advisor), Nel, Louis Hendrik (advisor), Markotter, Wanda (advisor).

Subjects/Keywords: African lyssaviruses; Polymerase chain reaction (PCR); Pcr-based methods; UCTD

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

author], [. (2010). Development of PCR-based methods for detection of African lyssaviruses . (Masters Thesis). University of Pretoria. Retrieved from http://upetd.up.ac.za/thesis/available/etd-10082010-143715/

Chicago Manual of Style (16^th Edition):

author], [No. “Development of PCR-based methods for detection of African lyssaviruses .” 2010. Masters Thesis, University of Pretoria. Accessed April 10, 2021. http://upetd.up.ac.za/thesis/available/etd-10082010-143715/.

MLA Handbook (7^th Edition):

author], [No. “Development of PCR-based methods for detection of African lyssaviruses .” 2010. Web. 10 Apr 2021.

Vancouver:

author] [. Development of PCR-based methods for detection of African lyssaviruses . [Internet] [Masters thesis]. University of Pretoria; 2010. [cited 2021 Apr 10]. Available from: http://upetd.up.ac.za/thesis/available/etd-10082010-143715/.

Council of Science Editors:

author] [. Development of PCR-based methods for detection of African lyssaviruses . [Masters Thesis]. University of Pretoria; 2010. Available from: http://upetd.up.ac.za/thesis/available/etd-10082010-143715/

Universidade Federal de Mato Grosso do Sul

20. Cominetti, Marlon Cezar. Ecoepidemiologia da Entomofauna Triatominae (Hemiptera: Reduviidae) e distribuição populacional das discretas unidades de tipagem (DTU) de Trypanosoma cruzi em Mato Grosso do Sul, Brasil .

Degree: 2015, Universidade Federal de Mato Grosso do Sul

URL: http://repositorio.cbc.ufms.br:8080/jspui/handle/123456789/2914

► Este trabalho decorre do estudo ecoepidemiologico da entomofauna Triatominae (Hemiptera: Reduviidae) e distribuição populacional das unidades discretas de tipagem (DTU) de Trypanosoma cruzi em Mato… (more)

▼ Este trabalho decorre do estudo ecoepidemiologico da entomofauna Triatominae (Hemiptera: Reduviidae) e distribuição populacional das unidades discretas de tipagem (DTU) de Trypanosoma cruzi em Mato Grosso do Sul, Brasil. Foi analisado o conteúdo intestinal de 515 triatomíneos, utilizando-se microscopia óptica e a Reação em Cadeia da Polimerase (PCR) para identificação de Trypanosoma cruzi. As amostras positivas para o parasito, proveniente de 12 triatomíneos, foram submetidas a uma segunda PCR, cujo alvo era o gene cromossômico TcSC5D, seguido da técnica de polimorfismo no comprimento de fragmentos de restrição (RFLP) com enzimas de restrição HpaI e SphI, clonagem e sequenciamento do amplificado. Em uma segunda etapa foram analisados, a partir de dados secundários, 14.178 espécimes de insetos vetores de T. cruzi capturados em 50 municípios de Mato Grosso do Sul em um período de nove anos e a influência de fatores climáticos, como temperatura, umidade relativa do ar e precipitação, sobre sua frequência. Observou-se que a PCR foi mais precisa na identificação do parasito que o exame de microscopia (19,6% e 11,3%, respectivamente). Foram identificadas três unidades discretas de tipagem (DTU): TcI (58,34%), TcII (33,33%) e TcBat/TcVII (8,33%). A espécie mais frequente no estado é Triatoma sordida (96,22%) e a maioria dos espécimes foi coletada em peridomicílio (85,06%). Verificou-se que a frequência desses insetos foi afetada por fatores climáticos como temperatura e umidade relativa do ar, sendo esta última a mais influente. Este trabalho aponta que os dados oficiais das taxas de infecção natural dos triatomíneos podem estar subestimados em todo o estado, apresenta a distribuição de três linhagens de T. cruzi, sendo que esse é o primeiro relato de infecção natural por TcBat/TcVII em triatomíneos e que as características climáticas ao longo do ano devem ser observadas pelas equipes de controle de vetores. Tais achados podem subsidiar programas de controle do vetor, propiciando a otimização dos recursos destinados às ações de prevenção e controle da transmissão vetorial de T. cruzi em Mato Grosso do Sul. Advisors/Committee Members: Andreotti, Renato (advisor).

Subjects/Keywords: Trypanosoma cruzi; Triatominae; Reação em Cadeia da Polimerase; Polymerase Chain Reaction

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Cominetti, M. C. (2015). Ecoepidemiologia da Entomofauna Triatominae (Hemiptera: Reduviidae) e distribuição populacional das discretas unidades de tipagem (DTU) de Trypanosoma cruzi em Mato Grosso do Sul, Brasil . (Doctoral Dissertation). Universidade Federal de Mato Grosso do Sul. Retrieved from http://repositorio.cbc.ufms.br:8080/jspui/handle/123456789/2914

Chicago Manual of Style (16^th Edition):

Cominetti, Marlon Cezar. “Ecoepidemiologia da Entomofauna Triatominae (Hemiptera: Reduviidae) e distribuição populacional das discretas unidades de tipagem (DTU) de Trypanosoma cruzi em Mato Grosso do Sul, Brasil .” 2015. Doctoral Dissertation, Universidade Federal de Mato Grosso do Sul. Accessed April 10, 2021. http://repositorio.cbc.ufms.br:8080/jspui/handle/123456789/2914.

MLA Handbook (7^th Edition):

Cominetti, Marlon Cezar. “Ecoepidemiologia da Entomofauna Triatominae (Hemiptera: Reduviidae) e distribuição populacional das discretas unidades de tipagem (DTU) de Trypanosoma cruzi em Mato Grosso do Sul, Brasil .” 2015. Web. 10 Apr 2021.

Vancouver:

Cominetti MC. Ecoepidemiologia da Entomofauna Triatominae (Hemiptera: Reduviidae) e distribuição populacional das discretas unidades de tipagem (DTU) de Trypanosoma cruzi em Mato Grosso do Sul, Brasil . [Internet] [Doctoral dissertation]. Universidade Federal de Mato Grosso do Sul; 2015. [cited 2021 Apr 10]. Available from: http://repositorio.cbc.ufms.br:8080/jspui/handle/123456789/2914.

Council of Science Editors:

Cominetti MC. Ecoepidemiologia da Entomofauna Triatominae (Hemiptera: Reduviidae) e distribuição populacional das discretas unidades de tipagem (DTU) de Trypanosoma cruzi em Mato Grosso do Sul, Brasil . [Doctoral Dissertation]. Universidade Federal de Mato Grosso do Sul; 2015. Available from: http://repositorio.cbc.ufms.br:8080/jspui/handle/123456789/2914

21. Λάζαρη, Σπυριδούλα. Polymerase chain reaction as a succesful biotechnological application. Ways we use PCR in the fields of bioinformatics forensics and genetics.

Degree: 2011, University of Patras

URL: http://nemertes.lis.upatras.gr/jspui/handle/10889/4575

►

Molecular genetics use molecular methods that amplify specific fragments of DNA. Today, the molecular techniques which were developed for amplification and detection of specific sequences… (more)

▼

Molecular genetics use molecular methods that amplify specific fragments of DNA. Today, the molecular techniques which were developed for amplification and detection of specific sequences of nucleic acids helped in a great deal to understand the structure of many diseases. Polymerase chain reaction or PCR is a technique that is used for isolation and amplification of a specific sequence of DNA. PCR is an in vitro method that exploits the in vivo procedure of replication of DNA. DNA polymerase is used to amplify a piece of DNA by in vitro enzymatic replication. As PCR progresses, the DNA thus generated is itself used as a template for replication. This sets in motion a chain reaction in which the DNA template is exponentially amplified. With PCR it is possible to amplify a single or few copies of a piece of DNA across several orders of magnitude, generating millions or more copies of the DNA piece. PCR was designed and presented by Dr Kary Mullis (1983).Today PCR has a great range of implementations related to 1) the cloning and the study of gene’s expression 2) the detection of mutations that are responsible for hereditary diseases 3) Criminology, Toxicology and Forensics.

H Μοριακή Γενετική χρησιμοποιεί μοριακές μεθόδους οι οποίες ενισχύουν συγκεκριμένα τμήματα DNA. Σήμερα, οι μοριακές τεχνικές οι οποίες αναπτύχθηκαν για ενίσχυση και ανίχνευση συγκεκριμένων ακολουθιών νουκλεϊνικών οξέων βοήθησαν σε μεγάλο βαθμό στην κατανόηση της δομής πολλών ασθενειών. Η αλυσιδωτή αντίδραση πολυμεράσης είναι μια τεχνική η οποία χρησιμοποιείται για απομόνωση και ενίσχυση μιας συγκεκριμένης ακολουθίας DNA. Η αλυσιδωτή αντίδραση της πολυμεράσης είναι μια διαδικασία που πραγματοποιείται σε ελεγχόμενες συνθήκες έξω απο ζωντανούς οργανισμούς, η οποία εκμεταλλεύεται την αντιγραφή του DNA που πραγματοποιείται μεσα στον ζωντανό οργανισμό. Όσο η PCR εξελίσσεται δημιουργούνται αντίγραφα DNA χρησιμοποιώντας ως πρότυπο το αρχικό DNA. Αυτό ενεργοποιεί μια διαδικασία αλυσιδωτής αντίδρασης ώπου το DNA αναπαράγεται εκθετικά. Με την PCR μπορείς να δημιουργήσεις εκατομμύρια αντίγραφα DNA από μία συγκεκριμένη ακολουθία. Η PCR σχεδιάστηκε και παρουσιάστηκε απο τον Dr Kary Mullis. Σήμερα η PCR έχει μέγαλο εύρος εφαρμογής σε πεδία σχετικά με τη μελέτη της γενετικής έκφρασης την ανίχνευση μεταλλάξεων οι οποίες είναι υπεύθυνες για κληρονομικές ασθένειες, Εγκληματολογία, Τοξικολογία και Ιατροδικαστική.

Advisors/Committee Members: Ματσόπουλος, Γεώργιος, Lazari, Spyridoula, Ματσόπουλος, Γεώργιος, Ουζούνογλου, Νικόλαος, Κουτσούρης, Διονύσιος.

Subjects/Keywords: Polymerase chain reaction (PCR); Genetics; 572.43; Αλυσιδωτή αντίδραση πολυμεράσης; Γενετική

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Λάζαρη, �. (2011). Polymerase chain reaction as a succesful biotechnological application. Ways we use PCR in the fields of bioinformatics forensics and genetics. (Masters Thesis). University of Patras. Retrieved from http://nemertes.lis.upatras.gr/jspui/handle/10889/4575

Chicago Manual of Style (16^th Edition):

Λάζαρη, Σπυριδούλα. “Polymerase chain reaction as a succesful biotechnological application. Ways we use PCR in the fields of bioinformatics forensics and genetics.” 2011. Masters Thesis, University of Patras. Accessed April 10, 2021. http://nemertes.lis.upatras.gr/jspui/handle/10889/4575.

MLA Handbook (7^th Edition):

Λάζαρη, Σπυριδούλα. “Polymerase chain reaction as a succesful biotechnological application. Ways we use PCR in the fields of bioinformatics forensics and genetics.” 2011. Web. 10 Apr 2021.

Vancouver:

Λάζαρη �. Polymerase chain reaction as a succesful biotechnological application. Ways we use PCR in the fields of bioinformatics forensics and genetics. [Internet] [Masters thesis]. University of Patras; 2011. [cited 2021 Apr 10]. Available from: http://nemertes.lis.upatras.gr/jspui/handle/10889/4575.

Council of Science Editors:

Λάζαρη �. Polymerase chain reaction as a succesful biotechnological application. Ways we use PCR in the fields of bioinformatics forensics and genetics. [Masters Thesis]. University of Patras; 2011. Available from: http://nemertes.lis.upatras.gr/jspui/handle/10889/4575

University of Johannesburg

22. Mienie, Charmain. Gene-specific PCR analysis of differential expression of the bean Chalcone Synthase multigene family.

Degree: 2012, University of Johannesburg

URL: http://hdl.handle.net/10210/6119

►

M.Sc.

A common feature of multi gene families is that their members are expressed in different ways in response to environmental and developmental signals. In… (more)

▼

M.Sc.

A common feature of multi gene families is that their members are expressed in different ways in response to environmental and developmental signals. In the present study the expression of a CHS multi gene family in bean (Phase°lus vulgari.), was studied. using a RT-PCR technique that focuses on the 3' divergent regions of the isogenes. Tissue-specific expression in roots, stems, leaves and the flowers of Phaseolus vulgaris, as well as in callus tissue, were investigated. Patterns and levels of gene expression were investigated after treatment with different elicitors as well as light. In most cases time and concentration studies were performed. The four CHS transcripts (CHS4. CHS1. CHS17 and CHS14) showed tissue-specific expression. The four CHS transcripts were differently expressed in the seven organs investigated: and different levels of activity were observed. The highest level of transcript expression for CHS14 and CHS4 could be observed in the roots, whereas relatively low levels were obtained in the leaves. stems and flowers of the green as well as etiolated seedlings. Higher levels of CHS were found in flower buds. High levels of all four transcripts were also found in callus. Elicitor treatment with structurally diverse abiotic agents showed induction of all four CHS mRNA transcripts. Concentration studies revealed high levels of CHS transcript levels. Elicitation with different concentrations of the elicitors: glutathione. mercuric chloride and sodium salicylate showed high levels of the CHS transcripts after exposure of 6 h to the different elicitation agents. The transcript levels increased significantly to levels above those observed in untreated (control) plants. The CHS transcripts showed higher levels of induction after elicitation with mercuric chloride (1 mM) relative to treatment with sodium salicvlate (10 rnM), suggesting differential regulation at the transcriptional level. The expression patterns observed with glutathione were very similarly to those induced by mercuric chloride. The kinetics of induction of all CHS transcripts. except for CHS1 were low at 2 and at 8 h postelicitation and maximal levels of transcript. although transiently induced, could be observed at 4 - 6 h. The use of 4 mM mercuric chloride did not give any induction. most probably because it was a lethal concentration. Etiolated and green bean seedlings, exposed to UV light. showed expression of all four CHS transcripts. In green leaves no significant differences in the induction kinetics between the different chs genes were observed. Three of the transcripts (CHS4. CHSI7. CHS14) accumulated rapidly (within ca. 3h). reaching a maximum after 6 h of irradiation. followed by a decline. CHS4 revealed a 18.2 fold induction. CHSI7 showed a 4.8 fold increase and CHS14 a 4 fold increase after 6 h of illumination in green leaves. In contrast. CHS1 showed a delay ed response which was still observable after 15 h. It was also demonstrated that CHS transcripts accumulated rapidly but transiently. Following illumination of…

Subjects/Keywords: Polymerase chain reaction; Common bean; Genetic engineering – Methods

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Mienie, C. (2012). Gene-specific PCR analysis of differential expression of the bean Chalcone Synthase multigene family. (Thesis). University of Johannesburg. Retrieved from http://hdl.handle.net/10210/6119

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Mienie, Charmain. “Gene-specific PCR analysis of differential expression of the bean Chalcone Synthase multigene family.” 2012. Thesis, University of Johannesburg. Accessed April 10, 2021. http://hdl.handle.net/10210/6119.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Mienie, Charmain. “Gene-specific PCR analysis of differential expression of the bean Chalcone Synthase multigene family.” 2012. Web. 10 Apr 2021.

Vancouver:

Mienie C. Gene-specific PCR analysis of differential expression of the bean Chalcone Synthase multigene family. [Internet] [Thesis]. University of Johannesburg; 2012. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10210/6119.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Mienie C. Gene-specific PCR analysis of differential expression of the bean Chalcone Synthase multigene family. [Thesis]. University of Johannesburg; 2012. Available from: http://hdl.handle.net/10210/6119

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Johannesburg

23. Jansen van Rensburg, Willem Sternberg. The development of a SCAR marker for the identification of the potato cultivars Astrid and Mnandi.

Degree: 2012, University of Johannesburg

URL: http://hdl.handle.net/10210/5524

►

M.Sc.

Mnandi and Astrid are two commercially important potato cultivars in South Africa. These two cultivars are closely related and are morphological virtually identical. It… (more)

▼

M.Sc.

Mnandi and Astrid are two commercially important potato cultivars in South Africa. These two cultivars are closely related and are morphological virtually identical. It is, however, necessary to be able to distinguish between these two cultivars, because each of these cultivars has certain desirable characteristics. It was decided to use DNA markers, since DNA markers are not influenced by the environment and the polymerase chain reaction (PCR) based DNA markers are relatively easy, cheap and fast. It was decided to develop a sequenced characterized amplified region (SCAR) due to the problems with the reproducibility of random amplified polymorphic DNA (RAPDs). SCARs are derived from RAPD fragments by using the sequence of a RAPD derived fragment to design a set of new longer primers (usually 20-24mer) which are less sensitive to PCR conditions. Ten commercial potato cultivars (Astrid, Mnandi. BP,, Buffelspoort, VanderPlank, Up-to-Date, Hoevelder, Hertha, Pimpernel and Agria) were used in this study. Commercially available RAPD primers (102) were evaluated to seek a polymorphism unique to either Mnandi or Astrid. Thirtyseven polymorphisms between Astrid and Mnandi were identified but only three were unique. The polymorphism obtained with OPH-15 was however, not reproducible. The polymorphisms obtained with UBC 509 and 582, corresponding to the presence in Mnandi of a 300 and 900 by fragment respectively, were reproducible. These two fragments, UBC 509 3" and UBC 582900, were cloned into the pMosBlue TA cloning vector and sequenced. The identity if the inserts in the recombinant plasmids were verified with PCR and Southern blotting. The sequences were used to develop two sets of SCAR primers, SCAR UBC 509 3" and SCAR UBC 582900 . The two SCAR primer pairs were then used in PCR reactions. The SCAR UBC 509 300 primer pair amplified a fragment of 230 by in both Astrid and Mnandi and a fragment of 260 by in Mnandi. The polymorphism is thus retained and SCAR UBC 509 3" can be used to distinguish between Astrid and Mnandi. The SCAR UBC 582' primer pair amplify a fragment of 500 by in both Astrid and Mnandi as well as some other longer fragments. It was not possible to regain a polymorphism by either elevating the annealing temperature or by digesting the amplification products with restriction enzymes. SCAR UBC 582' could thus not be used to distinguish between Astrid and Mnandi.

Subjects/Keywords: Potatoes – South Africa; Potatoes – Morphology; Polymerase chain reaction

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Jansen van Rensburg, W. S. (2012). The development of a SCAR marker for the identification of the potato cultivars Astrid and Mnandi. (Thesis). University of Johannesburg. Retrieved from http://hdl.handle.net/10210/5524

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Jansen van Rensburg, Willem Sternberg. “The development of a SCAR marker for the identification of the potato cultivars Astrid and Mnandi.” 2012. Thesis, University of Johannesburg. Accessed April 10, 2021. http://hdl.handle.net/10210/5524.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Jansen van Rensburg, Willem Sternberg. “The development of a SCAR marker for the identification of the potato cultivars Astrid and Mnandi.” 2012. Web. 10 Apr 2021.

Vancouver:

Jansen van Rensburg WS. The development of a SCAR marker for the identification of the potato cultivars Astrid and Mnandi. [Internet] [Thesis]. University of Johannesburg; 2012. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10210/5524.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Jansen van Rensburg WS. The development of a SCAR marker for the identification of the potato cultivars Astrid and Mnandi. [Thesis]. University of Johannesburg; 2012. Available from: http://hdl.handle.net/10210/5524

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Angelo State University

24. Sanders, Shanequa GeNey. DETERMINING THE PREVALENCE OF SALMONELLA.

Degree: Masters of Science, Animal Science, 2015, Angelo State University

URL: http://hdl.handle.net/2346.1/30414

► Experiment one’s objective was determining prevalence of Salmonella in small ruminants, their carcass and products. Experiment two’s objective was to validate a commercial PCR system… (more)

Subjects/Keywords: Salmonella; Polymerase Chain Reaction; small ruminants; Sheep; Goats

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Sanders, S. G. (2015). DETERMINING THE PREVALENCE OF SALMONELLA. (Masters Thesis). Angelo State University. Retrieved from http://hdl.handle.net/2346.1/30414

Chicago Manual of Style (16^th Edition):

Sanders, Shanequa GeNey. “DETERMINING THE PREVALENCE OF SALMONELLA.” 2015. Masters Thesis, Angelo State University. Accessed April 10, 2021. http://hdl.handle.net/2346.1/30414.

MLA Handbook (7^th Edition):

Sanders, Shanequa GeNey. “DETERMINING THE PREVALENCE OF SALMONELLA.” 2015. Web. 10 Apr 2021.

Vancouver:

Sanders SG. DETERMINING THE PREVALENCE OF SALMONELLA. [Internet] [Masters thesis]. Angelo State University; 2015. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/2346.1/30414.

Council of Science Editors:

Sanders SG. DETERMINING THE PREVALENCE OF SALMONELLA. [Masters Thesis]. Angelo State University; 2015. Available from: http://hdl.handle.net/2346.1/30414

University of Pretoria

25. Coertse, Jessica. Development of PCR-based methods for detection of African lyssaviruses.

Degree: Microbiology and Plant Pathology, 2010, University of Pretoria

URL: http://hdl.handle.net/2263/28545

► The etiological agent of rabies encephalitis belongs to the genus Lyssavirus in the Rhabdoviridae family. Lyssaviruses are negative sense, single stranded RNA viruses and cause… (more)

▼ The etiological agent of rabies encephalitis belongs to the genus Lyssavirus in the Rhabdoviridae family. Lyssaviruses are negative sense, single stranded RNA viruses and cause an estimated 55 000 human deaths per year with 44% of these deaths occurring in Africa (WHO, 2005). With intense research effort and increased sequence information it is becoming evident that the Lyssavirus genus is much more diverse than initially thought and therefore diagnostic methods need to be modified accordingly. The African continent sustains a diverse variety of lyssaviruses, however, most countries in Africa do not have active surveillance or necessary diagnostic tools and therefore rabies-related lyssaviruses are underreported. Previous studies have indicated that real-time PCR has improved sensitivity and rapidity over conventional molecular diagnostic methods with the added advantage of allowing accurate estimations of viral load in a wide variety of samples. Several realtime PCR assays have been developed; however, none were specifically aimed at detection of lyssaviruses present on the African continent. This study was therefore aimed at evaluating certain molecular diagnostic methods for the detection of African lyssaviruses. Furthermore, the application of real-time PCR for various fields in lyssavirus research i.e. diagnostics, surveillance and pathogenicity studies were evaluated. This study revealed two different hemi-nested PCR assays capable of detecting representatives of African lyssaviruses. A real-time PCR was developed that was successful for the detection of African lyssaviruses. In addition, a quantitative assay and internal control was successfully employed for confirming ante-mortem human rabies diagnosis as well as post-mortem animal rabies diagnosis in formalin fixed brain material. As such the real-time PCR assay developed in this study could therefore be routinely used for ante-mortem diagnosis and as a confirmatory test for post-mortem diagnosis. The ability of this assay to detect and quantify all currently known African lyssaviruses not only offers improved surveillance capacity, but offers unique potential as a sensitive tool to track virus movement in pathogenicity studies. These aspects are important in our search for a better understanding of the complex epidemiological and viral characteristics of African lyssaviruses. Copyright Advisors/Committee Members: Weyer, Jacqueline (advisor), Nel, Louis Hendrik (advisor), Markotter, Wanda (advisor).

Subjects/Keywords: African lyssaviruses; Polymerase chain reaction (PCR); Pcr-based methods; UCTD

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Coertse, J. (2010). Development of PCR-based methods for detection of African lyssaviruses. (Masters Thesis). University of Pretoria. Retrieved from http://hdl.handle.net/2263/28545

Chicago Manual of Style (16^th Edition):

Coertse, Jessica. “Development of PCR-based methods for detection of African lyssaviruses.” 2010. Masters Thesis, University of Pretoria. Accessed April 10, 2021. http://hdl.handle.net/2263/28545.

MLA Handbook (7^th Edition):

Coertse, Jessica. “Development of PCR-based methods for detection of African lyssaviruses.” 2010. Web. 10 Apr 2021.

Vancouver:

Coertse J. Development of PCR-based methods for detection of African lyssaviruses. [Internet] [Masters thesis]. University of Pretoria; 2010. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/2263/28545.

Council of Science Editors:

Coertse J. Development of PCR-based methods for detection of African lyssaviruses. [Masters Thesis]. University of Pretoria; 2010. Available from: http://hdl.handle.net/2263/28545

University of the Western Cape

26. Kasu, Mohaimin. Validation and application of a highly discriminating and rapid 10-locus Y-STR DNA profiling system .

Degree: 2019, University of the Western Cape

URL: http://hdl.handle.net/11394/6760

► DNA profiling the male specific region on the Y-chromosome is fundamental to forensic practise. Its recognised as a powerful analytical tool for investigation of sexual… (more)

Subjects/Keywords: DNA profiling; Forensic genetics; Polymerase Chain Reaction; Rapidly mutating; Sexual assault

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Kasu, M. (2019). Validation and application of a highly discriminating and rapid 10-locus Y-STR DNA profiling system . (Thesis). University of the Western Cape. Retrieved from http://hdl.handle.net/11394/6760

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Kasu, Mohaimin. “Validation and application of a highly discriminating and rapid 10-locus Y-STR DNA profiling system .” 2019. Thesis, University of the Western Cape. Accessed April 10, 2021. http://hdl.handle.net/11394/6760.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Kasu, Mohaimin. “Validation and application of a highly discriminating and rapid 10-locus Y-STR DNA profiling system .” 2019. Web. 10 Apr 2021.

Vancouver:

Kasu M. Validation and application of a highly discriminating and rapid 10-locus Y-STR DNA profiling system . [Internet] [Thesis]. University of the Western Cape; 2019. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/11394/6760.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Kasu M. Validation and application of a highly discriminating and rapid 10-locus Y-STR DNA profiling system . [Thesis]. University of the Western Cape; 2019. Available from: http://hdl.handle.net/11394/6760

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of the Western Cape

27. Mulder-Van Staden, Sune. Diode laser as an additional therapeutic measure in reducing red complex bacteria in chronic periodontitis .

Degree: 2016, University of the Western Cape

URL: http://hdl.handle.net/11394/5609

► This mini-thesis assessed whether a diode laser with a wavelength of 810 ± 10nm can be utilized as an adjunct to conventional management (i.e. scaling,… (more)

▼ This mini-thesis assessed whether a diode laser with a wavelength of 810 ± 10nm can be utilized as an adjunct to conventional management (i.e. scaling, root planing and polishing) of chronic periodontitis during initial phase therapy. Ethical approval and study registration (Reg no: 14/9/6) was finalized prior to commencement of the study. A split mouth randomised control trial was performed on 25 participants (who presented at the Oral Medicine and Periodontology Department of the University of the Western Cape) diagnosed with active, chronic periodontitis. In order to standardise the split mouth design the quadrants 1 & 4 were assessed together as a set and quadrants 2 & 3 were assessed as a set. A set of these quadrants were randomly assigned to either the test or control quadrants after conventional management was performed in all four quadrants. The base line bacterial colony collection (Micro-IDent®-11, Hain Lifescience GmbH, Nehren, Germany) and the clinical parameters were assessed prior to the commencement of conventional management and were reassessed at the 6 week re-evaluation visit. The set of test quadrants were treated with the diode laser as an adjunct to the preceding conventional management. The control quadrant only received the conventional management. Evaluation of the results demonstrated that the diode laser produced no statistical decrease in the bacterial parameters in the periodontal pockets and resulted in a statistical increase of C. showae (Cs) and T. denticola (Td). The clinical parameters resulted in no statistical difference for any clinical parameter, with the exception of the reduction in BOP that was statistically significant (p< 0,05) with the laser as an adjunct. It is the recommendation that within the limitations of this study, that the utilization of the diode laser (810 ± 10nm) as an adjunct at the initial visit had no statistical effect in the reduction of the bacterial parameters nor resulted in an overall improvement of the clinical parameters. Advisors/Committee Members: Holmes, H (advisor), Hille, J (advisor).

Subjects/Keywords: Polymerase chain reaction; Chronic periodontitis; Red complex bacteria; Periodontitis

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Mulder-Van Staden, S. (2016). Diode laser as an additional therapeutic measure in reducing red complex bacteria in chronic periodontitis . (Thesis). University of the Western Cape. Retrieved from http://hdl.handle.net/11394/5609

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Mulder-Van Staden, Sune. “Diode laser as an additional therapeutic measure in reducing red complex bacteria in chronic periodontitis .” 2016. Thesis, University of the Western Cape. Accessed April 10, 2021. http://hdl.handle.net/11394/5609.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Mulder-Van Staden, Sune. “Diode laser as an additional therapeutic measure in reducing red complex bacteria in chronic periodontitis .” 2016. Web. 10 Apr 2021.

Vancouver:

Mulder-Van Staden S. Diode laser as an additional therapeutic measure in reducing red complex bacteria in chronic periodontitis . [Internet] [Thesis]. University of the Western Cape; 2016. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/11394/5609.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Mulder-Van Staden S. Diode laser as an additional therapeutic measure in reducing red complex bacteria in chronic periodontitis . [Thesis]. University of the Western Cape; 2016. Available from: http://hdl.handle.net/11394/5609

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of the Western Cape

28. Tau, Tiroyamodimo. A forensic analysis of genetic variation in the Botswana population .

Degree: 2016, University of the Western Cape

URL: http://hdl.handle.net/11394/5657

► This thesis has been placed under a long term embargo. Forensic and population genetic parameters were investigated in the Botswana population using autosomal and Y-chromosome… (more)

▼ This thesis has been placed under a long term embargo. Forensic and population genetic parameters were investigated in the Botswana population using autosomal and Y-chromosome short tandem repeat markers. AmpFlSTR Profiler plus markers were used to investigate the genetic diversity and forensic parameters in 773 individuals from Botswana from the reference database of the Botswana Police. The levels of polymorphism found using the AmpFlSTR Profiler Plus markers showed that the nine loci that make up the AmpFlSTR Profiler Plus can differentiate individuals for forensic casework in the Botswana population. AmpFlSTR Identifiler autosomal STR markers were used to investigate the population structure according to ethno-linguistics and geography 990 individuals from Botswana that serve as a reference database for the Botswana Police. Using pairwise genetic distances (Fst), analysis of molecular variance (AMOVA), factorial correspondence analysis (FCA), and the unsupervised Bayesian clustering method found in STRUCTURE and the landscape genetics software TESS, ethno-linguistics were found to have a greater influence on population structure than geography. The patterns of population structure found using these markers highlight the need for regional reference databases that include both ethnolinguistic and geographic location information. These markers have important potential for bio-anthropological studies as well as for forensic applications. The 17 Y-chromosomal short tandem repeats found in AmpFlSTR Y-filer and a highly discriminatory Y-STR genotyping system (the Y-STR 10-plex developed in the Forensics DNA Laboratory at the University of the Western Cape) were analysed in 249 unrelated male individuals from Botswana. Rst, multi-dimensional scaling (MDS) and AMOVA were used to investigate population differentiation in Botswana. The discrimination capacity (DC) was found to be higher using the Y-STR 10-plex as compared to the 17 markers in the Y-filer genotyping system. No geographic regional or ethnic differentiation was observed between the Northern and Southern regions of Botswana using both marker systems. Regional and ethnic variation can be useful in forensic working hypotheses. Cluster analysis using the highly discriminatory Y-STR 10-plex haplotypes may provide information about ancestry and haplogroup information. Advisors/Committee Members: D’Amato, Maria Eugenia (advisor), Davison, Sean (advisor).

Subjects/Keywords: Polymerase Chain Reaction (PCR); Electrophoresis; Forensic genetics; Population genetics; Khoisan; Botswana

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Tau, T. (2016). A forensic analysis of genetic variation in the Botswana population . (Thesis). University of the Western Cape. Retrieved from http://hdl.handle.net/11394/5657

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Tau, Tiroyamodimo. “A forensic analysis of genetic variation in the Botswana population .” 2016. Thesis, University of the Western Cape. Accessed April 10, 2021. http://hdl.handle.net/11394/5657.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Tau, Tiroyamodimo. “A forensic analysis of genetic variation in the Botswana population .” 2016. Web. 10 Apr 2021.

Vancouver:

Tau T. A forensic analysis of genetic variation in the Botswana population . [Internet] [Thesis]. University of the Western Cape; 2016. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/11394/5657.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Tau T. A forensic analysis of genetic variation in the Botswana population . [Thesis]. University of the Western Cape; 2016. Available from: http://hdl.handle.net/11394/5657

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Waikato

29. Grout, Emily Katherine. Investigating molecular approaches for detection of pathogens in horses .

Degree: 2018, University of Waikato

URL: http://hdl.handle.net/10289/12094

► Infectious disease of equines has significant impacts on the New Zealand equine industry, causing avoidable morbidity and mortality. The correct identification of the aetiological agents… (more)

▼ Infectious disease of equines has significant impacts on the New Zealand equine industry, causing avoidable morbidity and mortality. The correct identification of the aetiological agents involved in such infections allows for prompt and accurate administration of an appropriate therapeutic strategy, reducing the economic impact associated with equine wastage. Traditional culturing methods are widely used for the diagnosis of infections in horses, however, conventional PCR offers a novel and rapid approach, through the targeting of unique genes for diagnosis of pathogens. PCR is an attractive alternative to microbial culturing, which can take anywhere between 24 hours to 1 week to determine the causative infectious agent, and in a number of cases the results are not as specific or as accurate as they need to be. It is the lack of sensitivity of the traditional culturing techniques that continues to contribute to the economic loss and welfare issues associated with equine infections. Initially, it was found that an SDS lysis extraction method worked best for extraction of DNA which was used to extract microbial DNA from various types of clinical samples. These were subsequently used as DNA templates within PCR alongside primers associated with up to 28 genes associated with common equine infections. From a total 146 clinical samples received for testing 66.4% returned a PCR positive for one or more microbial genera, whereas traditional bacteriological culturing produced an identification rate of 46%. In addition, PCR was significantly faster than culturing methods (6 hours vs 48 hours) and allowed more specific identification of the microbes present, which provides a significant advantage for infection management and increasing animal welfare. The results found from these experiments show rapid PCR identification holds promise as a diagnostic technique in the future, however, further development is required before this can be used as a primary diagnostic technique. Recommendations include further research into trends of infection in NZ equines and respectively increasing the diversity of PCR primers to represent the larger scope of microbial species. PCR diagnosis would also benefit from the standardisation of the DNA extraction methods used to allow consistency and increase throughput of various samples. Lastly, the simplification of the individual PCRs for each infectious agent could be developed into multiplex reactions to decrease time processing time and reduce human pipetting error. Advisors/Committee Members: Bird, Steve (advisor), Cursons, Raymond T (advisor).

Subjects/Keywords: Polymerase chain reaction; PCR; Molecular biology; Molecular genetics; Equine infectious disease

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Grout, E. K. (2018). Investigating molecular approaches for detection of pathogens in horses . (Masters Thesis). University of Waikato. Retrieved from http://hdl.handle.net/10289/12094

Chicago Manual of Style (16^th Edition):

Grout, Emily Katherine. “Investigating molecular approaches for detection of pathogens in horses .” 2018. Masters Thesis, University of Waikato. Accessed April 10, 2021. http://hdl.handle.net/10289/12094.

MLA Handbook (7^th Edition):

Grout, Emily Katherine. “Investigating molecular approaches for detection of pathogens in horses .” 2018. Web. 10 Apr 2021.

Vancouver:

Grout EK. Investigating molecular approaches for detection of pathogens in horses . [Internet] [Masters thesis]. University of Waikato; 2018. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10289/12094.

Council of Science Editors:

Grout EK. Investigating molecular approaches for detection of pathogens in horses . [Masters Thesis]. University of Waikato; 2018. Available from: http://hdl.handle.net/10289/12094

Kwame Nkrumah University of Science and Technology

30. Phillips, Richard O.; Frimpong, M; Sarpong-Duah, M; Beissner, M; Sarfo, F.S. Microscopy for Acid Fast Bacilli: A Useful but Neglected Tool in Routine Laboratory Diagnosis of Buruli Ulcer.

Degree: 2015, Kwame Nkrumah University of Science and Technology

URL: http://dspace.knust.edu.gh:8080/jspui/handle/123456789/11914

►

Background: Laboratory diagnosis of Buruli ulcer disease has become vital with the introduction of antibiotic treatment. Polymerase chain reaction (PCR) for the IS2404 repeat sequence… (more)

Subjects/Keywords: Mycobacterium ulcerans; Buruli ulcer; Ziehl-Neelsen staining; Polymerase chain reaction

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Phillips, Richard O.; Frimpong, M; Sarpong-Duah, M; Beissner, M; Sarfo, F. S. (2015). Microscopy for Acid Fast Bacilli: A Useful but Neglected Tool in Routine Laboratory Diagnosis of Buruli Ulcer. (Thesis). Kwame Nkrumah University of Science and Technology. Retrieved from http://dspace.knust.edu.gh:8080/jspui/handle/123456789/11914

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Phillips, Richard O.; Frimpong, M; Sarpong-Duah, M; Beissner, M; Sarfo, F S. “Microscopy for Acid Fast Bacilli: A Useful but Neglected Tool in Routine Laboratory Diagnosis of Buruli Ulcer.” 2015. Thesis, Kwame Nkrumah University of Science and Technology. Accessed April 10, 2021. http://dspace.knust.edu.gh:8080/jspui/handle/123456789/11914.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Phillips, Richard O.; Frimpong, M; Sarpong-Duah, M; Beissner, M; Sarfo, F S. “Microscopy for Acid Fast Bacilli: A Useful but Neglected Tool in Routine Laboratory Diagnosis of Buruli Ulcer.” 2015. Web. 10 Apr 2021.

Vancouver:

Phillips, Richard O.; Frimpong, M; Sarpong-Duah, M; Beissner, M; Sarfo FS. Microscopy for Acid Fast Bacilli: A Useful but Neglected Tool in Routine Laboratory Diagnosis of Buruli Ulcer. [Internet] [Thesis]. Kwame Nkrumah University of Science and Technology; 2015. [cited 2021 Apr 10]. Available from: http://dspace.knust.edu.gh:8080/jspui/handle/123456789/11914.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Phillips, Richard O.; Frimpong, M; Sarpong-Duah, M; Beissner, M; Sarfo FS. Microscopy for Acid Fast Bacilli: A Useful but Neglected Tool in Routine Laboratory Diagnosis of Buruli Ulcer. [Thesis]. Kwame Nkrumah University of Science and Technology; 2015. Available from: http://dspace.knust.edu.gh:8080/jspui/handle/123456789/11914

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Open Access Theses and Dissertations