Dissertations / Theses: 'Polymerase chain reaction (PCR)'

1

Feugeas, Olivier. "Pcr (polymerase chain reaction) et vih." Lille 2, 1990. http://www.theses.fr/1990LIL2M264.

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Lantz, Pär-G. "PCR-based detection of microorganisms in complex biological samples." Lund : Dept. of Applied Microbiology, Lund University, 1998. http://catalog.hathitrust.org/api/volumes/oclc/39178906.html.

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3

Ballagi-Pordány, András. "Application of polymerase chain reaction (PCR) in veterinary virology /." Uppsala : Sveriges lantbruksuniv, 1995. http://epsilon.slu.se/avh/1995/91-576-4997-9.gif.

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4

Erill, Sagalés Ivan. "High-speed Polymerase chain reaction in CMOS-compatible chips." Doctoral thesis, Universitat Autònoma de Barcelona, 2002. http://hdl.handle.net/10803/3031.

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En la última década del siglo XX, el campo de los microsistemas para análisis total (µ-TAS) y, más concretamente, el de los DNA-chips ha adquirido una importancia preponderante en el ámbito de los microsistemas. En gran parte, el creciente interés por estos dispositivos se debe a las substanciales mejoras que prometen: análisis más rápidos, baratos y automatizados, pero también es debido a la posibilidad de implementar técnicas analíticas antes impensables (e.g. chips de hibridación). En el caso particular de los DNA-chips, se han desarrollado prototipos funcionales para PCR, LCR, electroforesis en gel, di-electroforesis, hibridación y varias combinaciones de estas técnicas, al tiempo que los chips de hibridación masiva (mayoritariamente basados en arrayers) han llegado a convertirse en un éxito comercial. Aun así, y aunque se ha llevado a cabo mucho trabajo en estos años, es necesaria todavía mucha investigación para afrontar algunos de los principales retos de los DNA-chips.
En el transcurso de esta tesis doctoral, se ha llevado a cabo el desarrollo un proceso tecnológico común para la fabricación de DNA-chips multifunción (i.e. sistemas versátiles basados en PCR y electroforesis), poniendo un especial énfasis en la compatibilidad con los procesos CMOS estándar, a fin de conseguir desarrollar prototipos proto-industriales. Como demostrador de esta puesta a punto tecnológica, se han diseñado, fabricado y testado chips de PCR, y la PCR en chips ha sido optimizada con respecto a materiales de fabricación, metodologías de inserción/extracción, composición bioquímica de la mix de PCR, diferentes configuraciones de calentadores/sensores (Peltier/termopares vs. resistencias integradas) y la cinética de la reacción.
In the last decade of the twentieth century, the fields of µ-TAS and, more specifically, DNA-chips have acquired increasing importance in the microsystems arena. The main reason for this surge of interest lies in the advantages these new devices seek to bring forth: faster, cheaper and completely automated analyses, and also in the outbreak of novel analytical techniques (e.g. hybridization chips). In the particular case of DNA-chips, functional prototypes have been demonstrated for PCR, LCR, gel electrophoresis, di-electrophoresis, hybridization and various combinations of these techniques, whilst hybridization chips (mainly arrayer chips) have become a successful market application. But, even though a considerable amount of work has been carried out in these few years, much research is still required to address fundamental problems of DNA-chips.
In this doctoral work, a common-ground technological setup for the production of multifunction DNA-chips (i.e. PCR plus electrophoresis systems) has been laid down, placing strong emphasis in its compatibility with standard CMOS processes in order to produce proto-industrial prototypes. As a demonstrator of this technological setup, PCR-chips have been designed, manufactured and tested, and the chip PCR reaction has been optimized with respect to surface materials, insertion and extraction methods, biochemical mix composition, heater/sensor setups (Peltier/thermocouple vs. thin-film driven systems) and reaction kinetics.

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Aydin, Gamze. "Detection Of Genetically Modified Maize Via Polymerase Chain Reaction." Master's thesis, METU, 2004. http://etd.lib.metu.edu.tr/upload/3/12605495/index.pdf.

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In recent years, foods produced by genetic engineering technology have been on the world food market. The biosafety aspects, regulations, and labelling of these foods are still contentious issues in most countries. It is necessary to have approval for the use of GMOs in the production of food. Thus, detection and quantification of GMOs play crucial role for developing regulations on GM foods. In this study, raw and processed maize samples were analysed for genetic modification using a DNA based detection method, the Polymerase Chain Reaction. Ten raw food and 18 processed maize food including maize flour, starch, corn flakes, maize chips were collected from different markets located in different places in Turkey. The samples were examined for the presence of genetic elements located in the majority of transgenic crops such as NOS terminator, CaMV 35S promoter, kanamycin resistance (KanR) gene, using conventional PCR with oligonucleotide sets targeting to novel genes. Furthermore screening was conducted via Real-Time PCR assay for NOS terminator and 35S promoter. For confirming the presence of Bt11 maize lines event specific primers were utilised. Quantification of Bt11 maize lines were performed via Real-Time PCR. The result indicates that foreign genetic elements were found in all analysed raw material. In six out of 10 raw material, presence of Bt11 gene were identified. GMO detection was also possible for maize flour and starch, however in processed material as corn starch, corn flakes, corn chips and pop corn, transgenes were not detected.

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Allmann, Michael. "Applications of the polymerase chain reaction (PCR) in food chemistry /." [S.l.] : [s.n.], 1994. http://www.ub.unibe.ch/content/bibliotheken_sammlungen/sondersammlungen/dissen_bestellformular/index_ger.html.

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Duman, Zeynep. "Polymerase Chain Reaction (pcr) For Detection Of Borrelia Burgdorferi Sensu Lato." Master's thesis, METU, 2008. http://etd.lib.metu.edu.tr/upload/2/12609199/index.pdf.

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The present study aimed detection of a human pathogen B. bugdorferi sensu lato species in suspected Lyme borreliosis (LB) patients in Turkey by PCR analysis and supportive serologic tests. The 152 clinical samples (140 serum and blood, 10 cerebrospinal fluid (CSF), 1 synovial fluid, 1 skin biopsy specimens) from 140 patients sent from 22 different cities of Turkey to The Spirochetal Diseases Diagnosis Laboratory of Central Veterinary Control and Research Institute were analysed. Serum samples were subjected to ELISA with a commercial kit and all of the blood, CSF, synovial fluid and skin biopsy samples were examined by PCR. In PCR analysis two primer sets targeting the ospA gene located on the plasmid and ribosomal 23S rRNA gene of B. burgdorferi sensu lato were used. The results indicated that 32,1% (45 of 140) seropositivity was detectable by ELISA. Our results support that there is a risk of acquiring LB in different regions of Turkey. Although considerable positive detections were recorded using serologic tests,none of the specimens were positive in PCR analysis. Further studies on PCR based methods for detection of B. burgdorferi sensu lato in patients with a high clinical probability of LB apparently may require that the specimen should be taken in the early phases and before the administration of any therapeutic agent.

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Ros, Bascuñana Carlos. "Diagnostic application of the polymerase chain reaction (PCR) in veterinary microbiology /." Uppsala : Swedish Univ. of Agricultural Sciences (Sveriges lantbruksuniv.), 1997. http://epsilon.slu.se/avh/1997/91-576-5247-3.gif.

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Liao, Yu-Hua. "Polymerase chain reaction based cloning of acetate kinase in propionibacterium acidipropionici." The Ohio State University, 2004. http://rave.ohiolink.edu/etdc/view?acc_num=osu1072778140.

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Burns, Nigel. "Towards the absolute quantification of DNA by PCR." Thesis, Open University, 1999. http://oro.open.ac.uk/57922/.

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Amplification techniques such as the Polymerase Chain Reaction (PCR) are held to be largely qualitative procedures and are widely used as such. Since the efficiency of amplification is less than perfect, small changes in efficiency can yield dramatic differences in the final amount of product generated. Despite this unpredictability the exquisite sensitivity of PCR makes the demanding goal of absolute quantification highly desirable. Consequently, the use of this technique for the quantification of nucleic acids has increased at an exponential rate. However, the ability of PCR to accurately quantify absolute levels of DNA is still not universally accepted. The overall aim of this investigation was to determine the critical factors affecting the quantification of DNA using PCR and to use these findings to develop an assay for the absolute quantification of DNA in a model system. The novel work presented here illustrates the need for careful examination of sequencesfo r GC-rich domains which could give rise to stable secondary structures and reduce the efficiency of amplification by serving as termination sites. To determine the accuracy of competitive PCR, CE and IP-RP-HPLC were employed to quantify PCR- products. These two techniques provided valuable information on the identification and elimination of sources of error which led to improvements in speed, accuracy and precision, as well as ease of quantification by PCR. They also yielded information on the process of heteroduplex formation whilst simultaneously revealing assay limitations. Consequently, the on-line fluorescence monitoring of PCR was used as an alternative method for the quantification of Legionella pneumophila. This technique was highly reproducible however, mispriming and the subsequent amplification of non-specific PCR products limited the level of detection. The Y-end labelling of degraded DNA with DIG prevented short DNA fragments from mispriming (and consequently extending) allowing the amplification of DNA targets. Therefore, to reduce mispriming and hence improve assay sensitivity, this approach was adapted for the first time to produce 5'-degenerate, 3'- DIG-terminated competitive primer analogues. These analogues, coupled with the use of the LightcyclerTm, allowed the detection and absolute quantification of a single cell of Legionella pneumophila. This is the first time that this level of sensitivity has been achieved using this type of assay. This technique should provide a very rapid and sensitive alternative for quantification comparedt o the other,m oree xpensivete chnologiesa vailablea t present.

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Basten, Graham Paul. "Induction of phase II enzymes by isothiocyanates : an investigation using quantitative RT-PCR." Thesis, University of East Anglia, 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.393316.

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Balch, Signe Gyrite. "Cloning of novel macrophage-specific genes using differential-display PCR." Thesis, University of Oxford, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.312248.

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Lo, Terry. "Detection and Identification of Acinobacillus pleuropneumoniae serotype 5 by multiplex Polymerase Chain Reaction." Thesis, Virginia Tech, 1997. http://hdl.handle.net/10919/36902.

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Traditional serologic assays of Actinobacillus pleuropeumoniae often have problems with cross-reactivity. To avoid the complications of antibody-antigen reactions, a PCR assay was developed to detect Actinobacillus pleuropneumoniae and identify serotype 5 strains. Primers specific to the conserved capsular export region of A. pleuropneumoniae amplified a 0.7 kb DNA band in all strains with the exception of serotype 4. A second set of primers specific to the unique capsular biosynthesis region of serotype 5 amplified a unique 1.1 kb band for serotype 5 only. The sensitivity of this assay was determined to be less than 100 colony forming units. This PCR assay enables us to detect A. pleuronpeumoniae and definitively distinguishes serotype 5 strains from other serotyes.
Master of Science

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Bhattacharya, Shantanu. "A novel PCR based DNA microanalyzer system for detection of viral genome." Diss., Columbia, Mo. : University of Missouri-Columbia, 2006. http://hdl.handle.net/10355/4336.

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Thesis (Ph.D.)--University of Missouri-Columbia, 2006.
The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file viewed on (April 25, 2007) Vita. Includes bibliographical references.

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Liu, Tingting. "Electrokinetic Real-Time Polymerase Chain Reaction Toward Point-Of-Care Diagnosis." Diss., The University of Arizona, 2015. http://hdl.handle.net/10150/579083.

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Rapid diagnosis of infectious disease and timely initiation of proper clinical antibiotic treatment is the determinant in obtaining the optimal clinical outcomes and reducing emergences of multidrug-resistant organisms. In particular, acute infections require the detection to be accomplished in limited time with high sensitivity due to the low concentration of organisms causing the infections. Real-time Polymerase Chain Reaction can provide quantitative identification of specific genetic materials and has revolutionized clinical microbiology laboratory diagnosis. It is becoming a standard for infectious disease detection. However, most real-time PCR instruments on the market are bulky, fragile and costly due to their delicate optical components, which restricted their use to point-of-care application. Modern microfluidic and sensing technology provide a transition from benchtop real-time PCR to miniaturizable, robust, and portable real-time PCR devices to achieve rapid, low-cost, and efficient point-of-care diagnosis. In this work, an innovative electrokinetic PCR (EK-PCR) platform that combines AC electrothermal flow (ACEF) and Joule heating induced temperature gradient to implement thermal cycling for DNA amplification is discussed. In addition, in situ electrochemical sensing is incorporated in the EK-PCR chamber for real-time monitoring of the DNA concentration toward quantification of the initial copies of the DNA template. EK-PCR can improve the energy efficiency with minimized total thermal mass and remain high amplification efficiency. More importantly, it represents a highly integrated strategy for portable point-of-care devices.

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Halldén, Christer. "Characterization and use of a multiplex PCR-based system random amplified polymorphic DNA /." Lund : Lund University, 1998. http://catalog.hathitrust.org/api/volumes/oclc/68945134.html.

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Ahmad, M. Mursaleen Cheng Jianlin. "MUPrimer a tool for finding allele specific PCR-primers for homologous gene sequences /." Diss., Columbia, Mo. : University of Missouri--Columbia, 2009. http://hdl.handle.net/10355/6660.

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Title from PDF of title page (University of Missouri--Columbia, viewed on March 10, 2010). The entire thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file; a non-technical public abstract appears in the public.pdf file. Thesis advisor: Dr. Jianlin Cheng. Includes bibliographical references.

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Muwonge, Abubaker. "Detection Of Genetically Modified Potatoes By The Polymerase Chain Reaction." Master's thesis, METU, 2005. http://etd.lib.metu.edu.tr/upload/12605783/index.pdf.

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Quite a number of important crops have been genetically modified with genes for agronomically important traits, such as insect and viral resistance. As the numbers of genetically modified foods continue to increase on the market, the need for rapid development of GMO detection methods is indispensable. This study was carried out to detect if genetically modified potatoes exist on food market in Turkey. Thirty samples from different places were collected. Using a DNA based PCR method, potato samples were examined for the presence of 35S promoter, Nos terminator, neomycin phosphotransferase (nptII) genes, and synthetic cry3A gene which is the general transgene in all approved Newleaf transgenic potato lines. The experimental design of this study was to detect Newleaf insect resistant lines. In 11 samples at least one genetic element was detected. Sample R from Ankara has shown to be belonging to Newleaf insect resistant lines. Since 35S promoter was not detected in samples M3, 14 and F1, it is proposed that they are belonging to Newleaf virus and insect resistant lines (Newleaf plus or Newleaf Y). Although Nos terminator was not detected in samples H2, Z2 and D, cry3A fragments amplified in those samples have been verified that they are from the synthetic cry3A regions of Newleaf lines. The detected synthetic cry3A gene in GM potatoes was amplified by specific primers, which cannot amplify Bacillus thuringiensis tenebrionis natural cry3A gene. In addition, the authenticity of the synthetic cry3A PCR products were confirmed by both sequencing and restriction digestions. Our results showed that genetically modified Newleaf potatoes exist in food market in Turkey. Further studies by accredited laboratories are strongly recommended.

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Sonmezalp, C. Zeynep. "Detection Of Genetically Modified Insect Resistant Tomato Via Polymerase Chain Reaction." Master's thesis, METU, 2004. http://etd.lib.metu.edu.tr/upload/3/12605493/index.pdf.

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Tomato, which is one of the most important component of human diet, has been genetically modified to develop some properties like delayed ripening and insect resistance. In order to give a choice to the consumer, it is necessary to detect and label GM foods. This study was carried out to detect genetically modified tomato samples purchased from different food markets of Turkey. PCR method was used to detect genetically modified insect resistant tomatoes. The DNAs of collected samples were isolated according to CTAB DNA extraction protocol and the amplification capacity of isolated samples were checked with patatin specific control PCR. Screening tests of tomatoes were done by targeting 35S promoter, Nos terminator and NptII kanamycin resistance gene with four primer sets. It was aimed to detect Cry1A and Cry1Ac genes with three PCR systems, in order to identify insect resistant samples. From twenty-eight samples, twenty-two gave positive amplification signal in NptII specific PCR system and results were confirmed with sequence analysis. Additionally, we observed seventeen and ten DNA fragments with Cry1A-F/Cry1A-R and Cry1Ac-F/Cry1Ac-R primer sets respectively, it is necessary to confirm these results with DNA sequencing.

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Xu, Jingjing. "Magnetic particle based sensing platform for oligonucleotide and PCR amplicon detection /." View abstract or full-text, 2010. http://library.ust.hk/cgi/db/thesis.pl?CBME%202010%20XU.

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Cornett, Johanne Hazel. "The detection and quantification of genetically modified E. coli using PCR." Thesis, Birkbeck (University of London), 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.342184.

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Andrews, Daniel James. "Statistical models of PCR for quantification of target DNA by sequencing." Thesis, University of Cambridge, 2015. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.708581.

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Pallen, Mark John. "Detection and characterisation of diphtheria toxin genes and insertion sequences by PCR and other molecular techniques." Thesis, Queen Mary, University of London, 1993. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.245105.

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Mackay, Ian M. "Investigations into the utility of real-time PCR for the detection, quantitation and characterisation of clinically relevant viruses /." St. Lucia, Qld, 2003. http://adt.library.uq.edu.au/public/adt-QU20031120.155312/index.html.

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Lee, Thomas Ming Hung. "Sequence-specific electrochemical DNA detection and its implementation in integrated PCR-electrochemical microdevices /." View Abstract or Full-Text, 2003. http://library.ust.hk/cgi/db/thesis.pl?CENG%202003%20LEE.

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Thesis (Ph. D.)--Hong Kong University of Science and Technology, 2003.
Includes bibliographical references (leaves 155-177). Also available in electronic version. Access restricted to campus users.

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Duplàa, Cécile. "Analyse quantitative des produits de polymerase chain reaction utilisant l'incorporation de dUTP biotinylé." Bordeaux 2, 1993. http://www.theses.fr/1993BOR2P045.

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Gysling, Kevin. "Polymerase chain reaction restriction fragment length polymorphism identification of trebouxia lichen photosymbionts." Honors in the Major Thesis, University of Central Florida, 2009. http://digital.library.ucf.edu/cdm/ref/collection/ETH/id/1269.

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This item is only available in print in the UCF Libraries. If this is your Honors Thesis, you can help us make it available online for use by researchers around the world by following the instructions on the distribution consent form at http://library.ucf.edu/Systems/DigitalInitiatives/DigitalCollections/InternetDistributionConsentAgreementForm.pdf You may also contact the project coordinator, Kerri Bottorff, at kerri.bottorff@ucf.edu for more information.
Bachelors
Medicine
Molecular Biology and Microbiology

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胡婉晶 and Yuen-ching Wu. "Set up and validation of an automated PCR diagnostic and surveillance platform for influenza." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2009. http://hub.hku.hk/bib/B42905102.

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Wu, Yuen-ching. "Set up and validation of an automated PCR diagnostic and surveillance platform for influenza." Click to view the E-thesis via HKUTO, 2009. http://sunzi.lib.hku.hk/hkuto/record/B42905102.

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Gunson, Rory Niall. "The development, implementation and evaluation of a real-time PCR-based diagnostic service for viral causes of infectious intestinal disease." Connect to e-thesis, 2007. http://theses.gla.ac.uk/108/.

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Thesis (Ph.D.) - University of Glasgow, 2007.
Ph.D. thesis submitted to the Division of Community Based Sciences, Faculty of Medicine, University of Glasgow, 2007. Includes bibliographical references. Print version also available.

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Kumar, Sumeet Ph D. Massachusetts Institute of Technology. "Model microfluidic platform prototyping : design and fabrication of a Polymerase Chain Reaction (PCR) chip." Thesis, Massachusetts Institute of Technology, 2009. http://hdl.handle.net/1721.1/50572.

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Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2009.
Includes bibliographical references (p. 96-99).
Polymerase Chain Reaction (PCR) is a molecular biology method for the in vitro amplification of nucleic acid molecules, which has wide applications in the areas of genetics, medicine and biochemistry. MEMS technology offers several advantages for the miniaturization of biological protocols like PCR, including decreased amplification time, reduced reagent consumption, disposability, target specific amplification, and functional integration. The typical three step PCR cycle consists of heating the sample to 90-95 °C to denature the double-stranded DNA complex, cooling down to 55-60 °C to anneal the specific primers to the single stranded DNA, and finally increasing the temperature to 70-75 °C for extension of the primers with thermostable DNA polymerase. The temperature sensitivity of the reaction requires precise temperature control and proper thermal isolation of the three temperature zones. In this thesis, the design of a continuous flow PCR microfluidic platform consisting of a monolithic polydimethylsiloxane (PDMS) microfluidic chip assembled on top of a thin film patterned glass base heating unit is presented. A detailed thermo-fluidic model of the device is presented to predict the performance and efficacy of the proposed design. Numerical simulations are carried out to find the temperature distribution in the device and show the suitability of the design in meeting target temperature profile. Subsequently, simulation results are substantiated with experimental results of infrared and thermocouple temperature measurement on the device.
(cont.) An instrumented microfluidic platform was developed and experiments were carried out to investigate amplification efficiency. Different vapor barrier mechanisms and channel coatings were explored for minimizing sample loss. The research presented is an effort towards developing miniaturized, cost-effective, portable platform capable of replacing conventional thermocyclers.
by Sumeet Kumar.
S.M.

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Lefrère, Jean-Jacques. "Utilisation de la reaction d'amplification en chaine ou polymerase chain reaction (pcr) dans l'infection par le vih et ses applications en transfusion sanguine." Paris 6, 1993. http://www.theses.fr/1993PA066732.

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La technique de reaction d'amplification en chaine, ou polymerase chain reaction (pcr), permet une detection particulierement sensible des sequences virale. Elle s'est imposee comme outil de connaissance physiopathologique de l'histoire naturelle de l'infection par le virus de l'immunodeficience humaine (vih) et de l'efficacite des therapeutiques antivirales. Nous avons applique cette technique de biologie moleculaire dans differents contextes touchant la securite transfusionnelle vis-a-vis du vih et le developpement d'une therapeutique transfusionnelle du sida: 1) l'application de la pcr de l'adn du vih chez les sujets adultes appartenant a un groupe a risque, afin de valider la signification diagnostique des tests serologiques usuels, l'existence de sujets durablement seronegatifs et neanmoins infectes ayant pu remettre en cause les principes de securite transfusionnelle. 2) la constitution et la coordination d'un controle de qualite de la pcr de l'adn du vih, entrepris a l'echelon national, afin de faire de cette technique diagnostique un outil valide et partiellement standardise. 3) la mise au point d'une technique de quantification de l'adn du vih et son application dans l'etude de l'histoire naturelle de l'infection et dans la predictivite de la tendance evolutive vers la maladie chez les sujets seropositifs asymptomatiques. 4) la mise au point d'une technique de pcr permettant la mise en evidence de l'arn du vih. 5) l'etude de l'efficacite biologique d'une therapeutique transfusionnelle nouvelle du sida (immunotherapie passive), a l'aide de la mesure de la charge virale en adn par pcr quantitative et de la detection de l'arn du vih par pcr

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Glare, Eric M. 1965. "The application of competitive PCR technology to asthma research." Monash University, Dept. of Medicine, 2001. http://arrow.monash.edu.au/hdl/1959.1/7975.

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Kaddouche, Frédérique. "Utilisation de l'amplification génique (polymerase chain reaction) pour un criblage rapide des hémocultures." Montpellier 1, 1992. http://www.theses.fr/1992MON11230.

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Mohammed, Saleem. "Molecular studies on the protozoan parasite Toxoplasma gondii." Thesis, University of Reading, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.262211.

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THIRION, VINCENT. "Evaluation de la polymerase chain reaction (pcr) dans le diagnostic des infections a parvovirus b19." Lille 2, 1994. http://www.theses.fr/1994LIL2M237.

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Allery, Annie. "Amplification du gène ABL par PCR in situ dans des suspensions cellulaires (contribution technique)." Bordeaux 2, 1995. http://www.theses.fr/1995BOR2P080.

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Cabral, Paula Brito e. "Perfil de marcadores sorolÃgicos, salivares e moleculares de Mycobacterium leprae entre contatos intradomiciliares de pacientes com hansenÃase." Universidade Federal do CearÃ, 2012. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=8470.

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Os contatos de pacientes com hansenÃase Ã um grupo de alto risco de desenvolver a doenÃa, logo a precocidade no diagnÃstico Ã uma importante ferramenta para o controle da enfermidade. NÃs investigamos a presenÃa de DNA de Mycobacterium leprae, atravÃs da ReaÃÃo em Cadeia Polimerase (PCR), na mucosa nasal e anticorpos anti-PGL1 sÃricos, IgG/IgM, e salivares, IgA/IgM, por meio do ensaio imunoenzimÃtico ligado Ã fase sÃlida (ELISA), em contatos intradomiciliares de pacientes com hansenÃase (n=135) e seus casos Ãndices. O estudo foi realizado em duas cidades endÃmicas para a doenÃa no CearÃ, Crato e MaracanaÃ. Uma boa correlaÃÃo entre os isotipos IgA e IgM salivares e IgG sÃrico foi observada quando comparou-se os contatos intradomiciliares e seus casos Ãndices (p<0,01, p<0,005 e p<0.0001, respectivamente). Entretanto, essa relaÃÃo nÃo foi observada para IgM sÃrico anti-PGL1 (p>0,05). Observou-se alta frequÃncia de IgM sÃrico anti-PGL1 no grupo de contatos negativos para IgG sÃrico anti-PGL1 (71/121, 58,6%). Dentre os contatos positivos para IgG sÃrico anti-PGL1, o nÃvel de anticorpos sÃricos IgM anti-PGL1 positivos tambÃm foi alto (10/14, 71,4%). Em relaÃÃo aos anticorpos salivares, contatos de pacientes com a forma clÃnica PB mostraram boa correlaÃÃo entre IgA e IgM (r = 0.54, p <0.0001); o mesmo foi verificado em contatos de pacientes com a forma clÃnica MB (r = 0.61, p <0.0001). Foi observado que dos contatos positivos para os anticorpos salivares anti-PGL1, 53,3% obtiveram nÃveis de anticorpos sÃricos negativos. Por outro lado, dos contatos que foram negativos para os anticorpos salivares, 71,1% obtiveram nÃveis de anticorpos sÃricos positivos. O DNA de Mycobacterium leprae foi encontrado em swabs nasais de 9 contatos de pacientes portadores da forma clÃnica MB da hansenÃase (10,6%) e em 3 contatos de pacientes portadores da forma clÃnica PB (6.0%). NÃs concluÃmos que anÃlises quantitativas de anticorpos sÃricos e salivares anti-PGL1 em contatos de pacientes com hansenÃase sÃo necessÃrias para montar estratÃgias de levantamento de infecÃÃes subclÃnicas da hansenÃase, a fim de prevenir o desenvolvimento da doenÃa.
Leprosy household contacts are group at high risk of developing the disease. We investigated the presence M. leprae DNA in nasal mucosa and anti-PGL1 serum IgG/IgM and salivary IgA/IgM antibodies from leprosy household contacts (n=135) in two endemic regions, Ceara, Brazil. Good correlation between serum IgM and IgG isotypes was observed both in MB and in PB leprosy household contacts (r = 0.39, p <0.0001). However, their levels were much different (p <0.0001). Among the contacts positive for serum IgM, 74 (87%) were found to be negative for serum IgG. In respect to the salivary antibodies, PB leprosy household contacts showed correlation between IgA and IgM (r = 0.60, p <0.0001); the same was observed in MB leprosy contacts (r = 0.77, p <0.0001). It was observed that in 75.3% of the leprosy household contacts who were positive to serum anti-PGL1, their salivary antibodies were negative. On the other hand, 50% of the leprosy household contact who were negative to serum anti-PGL1 antibodies, their salivary antibodies were positive. M. leprae DNA was found in nasal swab in 9 MB household leprosy contacts (10.6%) and in 3 PB leprosy contacts (6.0%). We concluded that quantitative analysis of serum and salivary anti-PGL1 in leprosy contacts is necessary for mounting strategies to survey subclinical leprosy infections in order to prevent development of the disease.

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Bélanger, Elisabeth. "Genetic identification of the Lactobacillus species using PCR-based pepN sequences." Thesis, McGill University, 1998. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=21511.

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To improve the existing methods based on phenotype and/or genotype a new genotyping method was investigated to identify Lactobacillus species.
The method used the polymerase chain reaction (PCR) to amplify specific sequences of aminopeptidase (pepN) genes. The primers for the PCR reactions derived from a pepN sequence of Lactobacillus rhamnosus S93. PepN amplification products of 387 bp were obtained from forty three Lactobacillus strains and from some strains of Lactococcus (3), Streptococcus (2) and Bifidobactertium (5).
Restriction fragment length polymorphisms (RFLPs) and single-strand conformation polymorphisms (SSCP) methods were used to detect polymorphisms among amplified aminopeptidase DNA fragments from the different Lactobacillus strains.
The results of RFLPs after digestions with Sau3A I, Rsa I and Tru9 I confirmed that the PCR products were specific. According to the fingerprints generated, Lactobacillus species tested could be grouped in four.
SSCP allowed a good discrimination between different pepN PCR products of the same size. Some Lactobacillus strains, Lb. plantarum and Lb. rhamnosus showed the different ssDNA patterns. Though for many strains of Lactobacillus the SSCP patterns were similar, no general comparison can be made because all the samples were not loaded on the same SSCP polyacrylamide gel. The SSCP, PCR-based method can be easily modified to increase the rate of polymorphism detection.
This new genetic identification method is different from others because it uses specific pepN DNA sequences for each strain tested and it uses SSCP to detect the presence of polymorphisms. The method is also applicable to other genera of lactic acid bacteria.

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Schuchert, Jennifer Ann. "Detection of Actinobacillus Pleuropneumoniae and Identification of Serotypes 1, 2, and 8 by Multiplex Polymerase Chain Reaction." Thesis, Virginia Tech, 2002. http://hdl.handle.net/10919/34135.

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Traditional immunological assays used to serotype Actinobacillus pleuropneumoniae have been problematic due to cross- reactivity between serotypes, particularly serotypes 6 and 8. To avoid these serological cross-reactions, a multiplex PCR assay was developed to detect A. pleuropneumoniae and identify serotypes 1, 2, and 8. Primers specific to the conserved capsular polysaccharide export region of A. pleuropneumoniae serotype 5 amplified a 880 bp fragment in all serotypes excluding serotype 4 or a 489 bp DNA fragment in all serotypes including serotype 4. Primers specific to the capsular polysaccharide biosynthesis regions of A. pleuropneumoniae serotypes 1, 2, and 8 amplified a 1.6 kb, a 1.7 kb, and 970 bp fragment in the respective serotype. This PCR assay detects A. pleuropneumoniae and identifies serotypes 1, 2, and 8.
Master of Science

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Oshima, Yoko Pongrama Ramasoota. "Molecular subtyping of Aeromonas spp. using enterobacterial repetitive intergenic consensus (ERIC)-PCR /." Abstract, 2004. http://mulinet3.li.mahidol.ac.th/thesis/2547/cd363/4437638.pdf.

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LEE, SOOHYUN. "A NEW POLYMER LAB-ON-A-CHIP FOR POLYMERASE CHAIN REACTION (PCR) USING NON-CONTACT INFRARED THERMOCYCLES." University of Cincinnati / OhioLINK, 2005. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1123702712.

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Coertse, Jessica. "Development of PCR-based methods for detection of African lyssaviruses." Diss., University of Pretoria, 2010. http://hdl.handle.net/2263/28545.

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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
Dissertation (MSc)--University of Pretoria, 2010.
Microbiology and Plant Pathology
unrestricted

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Caine, Lesley-Anne. "Prevalence and antibiotic resistance determinants of Escherichia coli pathotypes obtained from raw milk in two farms from the Eastern Cape, South Africa: public health implications." Thesis, University of Fort Hare, 2013. http://hdl.handle.net/10353/d1015525.

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Milk quality continues to be a topic of intense debate in the dairy industry, medical and public health communities. Production of maximum quantities of high-quality milk is an important goal of every dairy operation. High-quality milk must contain a low number of somatic cells and low bacteria count, and must be free of human pathogens and antibiotic residues. The objective of this study was to determine the prevalence of E. coli in unpasteurized milk recovered from Middledrift and Fort Hare dairy. In this study 400 milk samples were collected from two commercial farms (Middledrift and Fort Hare) in the Eastern Cape, South Africa, 200 raw milk samples from each farm. Samples were cultured on violet red bile mug-agar (VRB-MUG Agar) and incubated at 37ºC for 24 hours and preliminary identified by Gram stain and catalase test. Isolates that were Gram negative and catalase positive were screened for a marker of E. coli uidA gene using PCR assays. Middledrift dairy farm had 50 (25%) E. coli isolated from raw milk and Fort Hare farm showed 37 (18.5%) E. coli present in the milk samples. The presence of E. coli found in the milk samples points to the fact that fecal contamination was unavoidable and traditional practices are likely to contribute to the contamination of the milk and proliferation of the microorganisms.

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Jacobs, Gwynneth. "Evaluation of insertion-deletion polymorphisms with the kit Qiagen Investigator® DIPplex for forensic application in South Africa." University of the Western Cape, 2015. http://hdl.handle.net/11394/4690.

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>Magister Scientiae - MSc
Insertion-deletion polymorphisms (indels) have been underutilized in forensic identification of individuals in comparison with single nucleotide polymorphisms (SNPs) and short tandem repeat (STRs) systems. The use of indels for the purpose of human identification is more advantageous than previously used methods as it combines desirable characteristics of both the SNPs and STRs i.e. low costs and simplistic typing methods as well as indels having small amplicons size, making them suitable for genotyping highly degraded DNA. Currently there is only one commercial kit available for the forensic community, the Investigator® DIPlex kit (Qiagen), which cover a total of 30 indel loci distributed over 19 autosomal chromosomes. The objective of this study was to evaluate the Qiagen Investigator® DIPplex kit for forensic application in South Africa. The kit‘s performance was evaluated by comparing different extraction methods; sensitivity, robustness and reproducibility were evaluated and forensic parameters (match probability, power of discrimination, polymorphism information content, power of exclusion and typical paternity index) were estimated based on population data generated from five South African populations (Afrikaner, Mixed Ancestry, Indian-Asian, Xhosa and Zulu). Population comparisons were performed using Fst-analysis, factorial component analysis as well as phylogenetic tree construction. DNA was extracted from buccal swabs and whole blood collected from a total of 512 individuals from the five South African population groups and genotyped using the Qiagen Investigator® DIPplex kit. Sanger DNA sequencing and sequence alignments confirmed the presence of a null allele at locus HLD97 which was present in high frequency in the Xhosa and Zulu populations. This observation was made in 14 individuals belonging to the Xhosa and Zulu populations. Null allele frequencies in all five South African populations were also estimated. Null alleles were estimated for all loci using analytical methods i.e. Charkraborty null allele estimator, Brookfield null allele estimators 1 and 2 and ML-NullFreq software program. The kit performed well in the laboratory, not requiring any additional reagents or instrumentation and successfully generating profiles with input DNA amounts as low as 0.2 ng/μL. Although well suited for forensic application, the Qiagen Investigator® DIPplex kit showed some drawbacks with regards to application on South African populations. The presence of a null allele at the HLD97 locus as well as indication of population substructure affects allele frequency estimates for the South African populations. Correction for population substructure as present within the South African populations should be considered using FST analysis and it is recommended that the HLD97 locus should be excluded from any kinship analysis performed on South African populations.

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Taniga, Velan. "Développement d’un module d’amplification par PCR avec suivi électrochimique pour la détection de bactérie." Thesis, Paris 6, 2015. http://www.theses.fr/2015PA066189.

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Nous avons développé un module de detection de bactérie base sur le suivi électrochimique d’une PCR. Le système a été realisé en Copolymère d’Oléfin Cyclique (COC). Il s’agit ici de présenter une nouvelle approche pour concevoir des systèmes de despistage d’infection nosocomiale en quelques heures. INTRODUCTION Avec le développement des flux migratoires de population et l’apparition de résistance aux antibiotiques, les infections nosocomiales sont devenues un enjeu de santé majeur. Les besoins en systèmes de diagnostique accessibles financièrement augmentent considérablement. L’approche qui consiste à intégrer entièrement une chaîne d’analyse commence à avoir les faveurs des systèmes de santé et en particulier de l’Organisation Mondiale de la Santé (OMS). Une compagnie nord américaine Cepheid a par ailleurs déjà développé un système automatisé de détection de germes basé sur l’identification de l’ADN par PCR. Cependant le coût élevé des technologies ralentit sa démocratisation malgré un apport considérable pour les tests cliniques de routine. Ici nous nous proposons une stratégie d’analyse qui permet de maîtriser les drastiquement les coûts de consommable. Tout d’abord tout le système est réalisé en COC, un thermoplastique qui peut être façonné à très grande échelle pour quelques dizines de centimes l’unité. [1]. Ensuite le système de détection, implique un suivi électrochimique qui remplace avantageusement les plateformes optiques traditionnellement coûteuses. Si la première cible de ce système est le dépistage de maladie nosocomiale, il apparaît aussi qu’il est transposable à d’autres domaines où la détection basée sur une PCR peut apporter des solutions. THEORIE L’échantillon, un écouvillon nasal est prélevé sur le patient. Ensuite les bactéries sont lysées grâce à l’utilisation de détergent. Une fois les parois bactériennes détruites, l’ADN est extrait sur phase solide par l’intermédiaire de billes magnétiques de type « charge switch » assemblées en colonnes [2]. Après lavage l’ADN est élué, puis amplifié par PCR en temps réel. Durant cette phase l’amplification de l’ADN est suivie grâce à l’utilisation d’un intercalant redox. Ce principe de détection fait l’objet d’un brevet [3] et nous présentons ici sa première implémentation dans un système de type laboratoire sur puce. Le complexe redox qui sert d’intercalant est détecté par voltammétrie à onde carrée (en anglais : Square Wave Voltammetry SQWV). Ce complexe interagit avec l’ADN double brin par intercalation pendant la phase d’élongation, ainsi il n’est plus disponible pour l’échange d’électrons avec les électrodes de détection. En conséquence, on observe une chute du niveau de courant mesuré pendant la SQWV qui correspond à l’augmentation de la quantité d’ADN. PRATIQUE De noveaux protocoles de fabrication de systèmes en thermoplastique ont été développés permettant de réaliser des système en COC robustes et étanches. L’intégration des électrodes sérigraphiées a été réalisée avec succès. Le desgin de la puce a été choisi afin de permettre la parallélisation d’expérience en vue des tests de sensibilité de détection. Afin de réaliser les cycles de chauffage nécessaire à l’amplification par PCR un système dédié, comprenant un bloc thermique, un système de régulation et une partie logicielle à été développé. La preuve de concept de l’efficacité du système a été démontré sur de l’ADN modèle : le Litmus. Il est possible de distinguer différentes concentration en ADN de départ
With the development of human mobility and antibiotics resistance, nosocomial infections have become a major health problem. The need for fast and efficient diagnosis systems, ,while affordable by the health care systems, is increasing. The “sample to result” integration allowed by microfludics is a strong asset in such developments. Cepheid developed an integrated system for germ genotyping based on real-time PCR, but due to expensive laser-induced fluorescence detection its cost still reduces its range of applications in routine clinics. Here, we propose a new strategy allowing further cost reduction. First, the whole analysis streamline is integrated in a single chip made of Cyclo Olefin Copolymer (COC) [1]. This material can be mass-processed by CD technology, to yield chips at a few cents a piece. Second, the detection involves an electrochemical method that alleviates need for optics. Methicillin Resistant Staphylococcus Aureus (MRSA) was chosen as the initial primary target for validation. The technologies developed will further be applied to other strains of Bacteria or other fields such as agriculture, food testing, GMOs or security. The sample, a nasal swab, undergoes a chemical lysis, and DNA is extracted by selective capture on magnetic beads self-assembled into dense microcolumns arrays, as presented at MicroTAS 2009 [2]. DNA is then eluted, amplified by PCR in real time in the chip, while detected in situ by a high sensitivity electrochemical detection catalyzed by an intercalating redox compound. This proprietary technology [3] is being implemented here for the first time in a lab on chip. New protocols were developed to make a robust and leakage-free COC microfluidic chip with integrated electrodes. COC is a biocompatible and solvent resistant thermoplastic material. The COC microfluidic chip consists of a substrate with a hot-embossed microchannel and screen printed carbon and silver electrodes. The chip is sealed by solvent bonding [4]. This provides simple and cost effective fabrication, directly upscalable to mass production PCR Thermal control is done externally with a customized thermocycler. A redox compound is introduced together with DNA sample enabling electrochemical detection through Square Wave Voltammetry (SQWV). This compound intercalates in double-stranded DNA during the extension step, reducing its mobility and redox activity, so the peak current decreases during amplification of the target DNA. A sensitivity below 1ng/µL suitable for MRSA detection was achieved. The data were obtained by SQWV measurements. There is a clear correlation between the increase of DNA concentration and the decrease of the peak current for the redox compound: PCR amplification was also performed inside the chip, demonstrating the compatibility of this platform with thermal cycling and biomolecular reactions. Real-time electrochemical measurement during cycling was not possible yet due to experimental setup geometrical constraints, but work is in progress and results will be presented at the conference. Future experiments will focus on the integration of the bacteria lysis and DNA extraction steps on the same chip

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王玲娜 and Ling-na Wang. "Direct detection of mycobacterium tuberculosis in clinical specimens by PCR-ELISA." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2001. http://hub.hku.hk/bib/B31969975.

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Wang, Ling-na. "Direct detection of mycobacterium tuberculosis in clinical specimens by PCR-ELISA." Hong Kong : University of Hong Kong, 2001. http://sunzi.lib.hku.hk/hkuto/record.jsp?B23476564.

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Chen, Hong. "Identification of novel Wilms' tumor related genes by using differential display polymerase chain reaction, DD-PCR." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1997. http://www.collectionscanada.ca/obj/s4/f2/dsk3/ftp04/mq22583.pdf.

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Vögtlin, Andrea. "Use of polymerase chain reaction (PCR) for the detection of vaccine contamination by infectious laryngotracheitis virus /." [S.l.] : [s.n.], 1998. http://www.ub.unibe.ch/content/bibliotheken_sammlungen/sondersammlungen/dissen_bestellformular/index_ger.html.

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Dissertations / Theses on the topic 'Polymerase chain reaction (PCR)'

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