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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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Verhaegen, Monique Elise. "Novel approaches in quantitative polymerase chain reaction." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1998. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape10/PQDD_0021/MQ52489.pdf.
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Chiou, Jeffrey Tsungshuan. "A novel capillary polymerase chain reaction machine." Thesis, Massachusetts Institute of Technology, 2001. http://hdl.handle.net/1721.1/8864.
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Thesis (Ph.D.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2001.Includes bibliographical references (p. 254-268).
I built a novel prototype capillary polymerase chain reaction machine. The purpose was to perform a single reaction as fast as possible with a reaction volume - 100 nl. The PCR mix is in the form of a 1 /1 droplet that moves between three heat zones inside of a 1 mm I.D. capillary filled with mineral oil via pneumatic actuation. A laser beam waveguides down the capillary until it strikes the drop, at which point it scatters. The scatter is picked up by a series of photodiodes to provide position feedback. Due to the efficient heat transfer arrangement, the drop can transition between different temperature steps in -2 seconds, which includes both drop motion and temperature equilibration. It was extensively tested in both 10-cycle and 30-cycle PCR, including nearly 200 successful 30-cycle runs. The 30-cycle PCR was typically 74% (as high as 78%) efficient, and took only 23 minutes. This compares well with existing machines in the literature.
by Jeffrey Tsungshuan Chiou.
Ph.D.
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Clackson, Timothy Piers. "Antibody engineering using the polymerase chain reaction." Thesis, University of Cambridge, 1991. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.316695.
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Linley, M. "The detection of polymerase inhibiting lesions using the polymerase arrest polymerase chain reaction assay." Thesis, Swansea University, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.637924.
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There is a constant need to determine the genotoxic potential of the agents to which the human population is exposed. The stringent testing of new products is legislatively controlled and dependent on the accumulation of sufficient scientific data to allow an analysis of the risk. It is important to predetermine any risks in the workplace prior to the presentation of disease and to provide factual public information on personal exposure e.g. the risks associated with UV light. Various experimental assays have been developed to assess the genotoxicity, mutagenicity and mcarcinogenicity of given physical and chemical agents. The Polymerase Arrest- Polymerase Chain Reaction (PA-PCR) assay was employed to investigate the genotoxic effects (DNA adducts, DAN strand breaks and DNA crosslinking) of various physical and chemical agents on naked isolated DNA. The assay was modified to provide two adapted methods, which increased the sensitivity of the assay to report DNA damage at significantly lowered exposure levels. The ability of the PA-PCR assay to perform as an initial screening process for genotoxic activity was assessed and determined.
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Nebbali, M. "Human gene mapping using the polymerase chain reaction." Thesis, University of Nottingham, 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.317395.
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Borges, Pinto Lais Izabel. "Alu-polymerase chain reaction genomic fingerprinting in neuroblastoma." Thesis, University of Newcastle Upon Tyne, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.366679.
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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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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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Woolford, Alison Jane. "Use of polymerase chain reaction for detection of mycobacteria." Thesis, University of Surrey, 1995. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.308497.
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Gurram, Neil (Neil K. ). "A mathematical model of polymerase chain reaction induced stutter." Thesis, Massachusetts Institute of Technology, 2016. http://hdl.handle.net/1721.1/106012.
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Thesis: M. Eng., Massachusetts Institute of Technology, Department of Electrical Engineering and Computer Science, 2016.This electronic version was submitted by the student author. The certified thesis is available in the Institute Archives and Special Collections.
Cataloged from student-submitted PDF version of thesis.
Includes bibliographical references (page 48).
This is a thesis on understanding stutter present in capillary electropherogram readouts as this methodology forms the basis of current DNA fingerprinting. The readouts come from taking samples of various initial template masses of DNA from different individuals, applying polymerase chain reaction (PCR) to the samples, and then running the amplified copies through capillary electrophoresis to produce a readout of peak heights corresponding to alleles on various loci. The alleles correspond to the number of repeats of microsatellites that are usually two to six base pairs in length called short tandem repeats (STRs); the number of repeats of various STRs defines a person's DNA fingerprint. This process introduces artifacts in measurement. Of particular interest in this thesis is stutter, the phenomenon where amplicons with fewer or greater number of STR repeats than the true allele count are generated as an artifact of the PCR. It is of interest to understand the source and nature for this stutter distribution for small starting masses, as it has ramifications on the ability to accurately determine a match between a DNA sample and a crime scene sample. Understanding the stutter distribution in this thesis is achieved through data analysis, probabilistic modeling, and statistics. We find that a mathematical model that combines stochastic effects from PCR with fluorescent noise explains the most significant features of the observed phenomena.
by Neil Gurram.
M. Eng.
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Lewis, Monte. "Thermal cycling design alternatives for the polymerase chain reaction." College Park, Md. : University of Maryland, 2005. http://hdl.handle.net/1903/3061.
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Thesis (M.S.) -- University of Maryland, College Park, 2005.Thesis research directed by: Dept. of Mechanical Engineering. Title from t.p. of PDF. Includes bibliographical references. Published by UMI Dissertation Services, Ann Arbor, Mich. Also available in paper.
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Way, Jaw-Shiow Chu. "Specific detection of Salmonella by multiplex polymerase chain reaction." Diss., The University of Arizona, 1993. http://hdl.handle.net/10150/186207.
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This study evaluates the use of multiplex polymerase chain reaction (PCR) technology for detection of Salmonella species in pure cultures and also in environmental samples. Three sets of oligonucleotide primers were used in the PCR assay: PhoP primers specific to a 299 bp region in the phoP/phoQ loci of coliform pathogenic bacteria such as Salmonella, Shigella, E. coli and Citrobacter species, served as a presumptive indicator of enteric bacteria. Subsequently Hin and H-1i primers, which targeted a 236 bp region of the hin/H2 gene, and a 173 bp region of the H-1-i flagellar gene in Salmonella typhimurium respectively, were used for specific detection of salmonellae. Specificity and sensitivity of PCR amplified products were evaluated by ethidium bromide (EtBr) staining or gene probe analysis. Under optimal PCR conditions described in this study, Salmonella species can be specifically detected by these 3 primer sets. The sensitivity of detection in terms of whole cells was a 10⁻⁵ dilution (approximately 10³ cells) of boiled late log phase cultured cells (detection by EtBr) after 25 cycles of PCR and 10⁻⁸ dilution (approximately 10° cells) after a 50 cycle 'double PCR' protocol. A similar level of sensitivity was observed using a gene probe analysis (³²P end-labeling) to detect the PCR amplified products. The multiplex PCR products observed from known environmental isolates allowed identification of several isolates as Salmonella species. These results agreed with conventional analyses. The efficacy of this PCR technology for environmental applications was also evaluated by testing the primers on soil and environmental water samples. All unseeded soil samples showed no PCR amplified products when detected by EtBr staining after 25 and 50 cycles PCR. Only one pond surface water sample resulted in a positive PCR result. This water sample was also positive when tested by conventional methodologies. Seeded soil and well water samples all resulted in PCR products being observed from 25 or 50 cycles. These results indicate that the primers have the potential to specifically detect Salmonella species in environmental samples.
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Sikorsky, Jan A. "Effect of DNA base modification on polymerase chain reaction efficiency and fidelity." Huntington, WV : [Marshall University Libraries], 2005. http://www.marshall.edu/etd/descript.asp?ref=554.
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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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White, Adam. "Development and application of microfluidic single-cell polymerase chain reaction." Thesis, University of British Columbia, 2015. http://hdl.handle.net/2429/55734.
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Methods for single-cell analysis are critical to revealing cell-to-cell variability in biological systems, such as during development or onset of disease, where the characteristics of heterogeneity and minority cell populations are obscured by population-averaged measurements. Analysis of individual cells has been limited due to challenges associated with small amounts of starting material, combined with the cost and throughput required to examine large numbers of cells. Microfluidic approaches are well suited to single-cell analysis, providing increased sensitivity, economy of scale, and automation. This thesis presents the development and application of microfluidic technology for single cell gene expression analysis. The foundational contribution of this work is an integrated microfluidic device capable of performing high-precision RT-qPCR measurements of gene expression from hundreds of single cells per run. This device executes all steps of single cell processing including cell capture, cell lysis, reverse transcription, and quantitative PCR. This device is further expanded upon by integrating the single cell and nucleic acid processing capabilities with final measurement of cDNA by high-density digital PCR. The direct quantification of single molecules by digital PCR has advantages over RT-qPCR in the measurement of low abundance transcripts, as well as obviating the need for relative abundance measurements or calibration standards. This technology is demonstrated in over 5,000 individual cell measurements of mRNA, microRNA, and single nucleotide variant detection in a variety of cell types. Finally, this technology is applied to study the performance of lipid nanoparticles in delivery of RNA, and manipulation of gene expression in cells. The microfluidic integration of cell and nucleic acid processing established in this thesis permits analysis of hundreds of single cells in parallel, while improving work flow and reducing technical variation compared to samples prepared in microliter volumes. Ultimately, this advances the tools available for precisely measuring transcripts in single cells, and has application in research and clinical settings.Science, Faculty of
Graduate
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Tsang, Tsui-ying Stella, and 曾璀瑩. "Application of quantitative polymerase chain reaction in the diagnosisof thalassaemia." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2006. http://hub.hku.hk/bib/B45010948.
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Gamma, Torres Rafael Enrique. "Application of polymerase chain reaction to rhinovirus detection and analysis." Thesis, University of Essex, 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.293589.
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Koc, Yasemin. "Optimization of continuous flow polymerase chain reaction with microfluidic reactors." Thesis, Imperial College London, 2008. http://hdl.handle.net/10044/1/8184.
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The polymerase chain reaction (PCR) is an enzyme catalyzed technique, used to amplify the number of copies of a specific region of DNA. This technique can be used to identify, with high-probability, disease-causing viruses and/or bacteria, the identity of a deceased person, or a criminal suspect. Even though PCR has had a tremendous impact in clinical diagnostics, medical sciences and forensics, the technique presents several drawbacks. For example, the costs associated with each reaction are high and the reaction is prone to contamination due to its inherent efficiency and high sensitivity. By employing microfluidic systems to perform PCR these advantages can be circumvented. This thesis addresses implementation issues that adversely affect PCR in microdevices and aims to improve the efficiency of the reaction by introducing novel materials and methods to existing protocols. Molecule-surface-interactions and temperature control/determination are the main focus within this work. Microchannels and microreactors are characterized by extremely high surface-to-volume ratios. This dictates that surfaces play a dominant role in defining the efficiency of PCR (and other synthetic processes) through increased molecule-surface interactions. In a multicomponent reaction system where the concentration of several components needs to be maintained the situation is particularly complicated. For example, inhibition of PCR is commonly observed due to polymerase adsorption on channel walls. Within this work a number of different surface treatments have been investigated with a view to minimizing adsorption effects on microfluidic channels. In addition, novel studies introducing the use of superhydrophobic coatings on microfluidic channels are presented. Specifically superhydrophobic surfaces exhibiting contact angles in excess of 1500 have been created by growing copper oxide and zinc oxide nanoneedles and silica-sol gel micropores on microfluidic channels. Such surfaces utilize additional surface roughness to promote hydrophobicity. Aqueous solutions in contact with superhydrophobic surfaces are suspended by bridging-type wetting, and therefore the fraction of the surface in contact with the aqueous layer is significantly lower than for a flat surface. An additional difficulty associated with PCR on microscale is the detennination and control of temperature. When perfonning PCR, the ability to accurately control system temperatures is especially important since both primer annealing to single-stranded DNA and the catalytic extension of this primer to form the complementary strand will only proceed in an efficient manner within relatively narrow temperature ranges. It is therefore imperative to be able to accurately monitor the temperature distributions in such microfluidic channels. In this thesis, fluorescence lifetime imaging (FLIM) is used as a novel method to directly quantify temperature within microchannel environments. The approach, which includes the use of multiphoton excitation to achieve optical sectioning, allows for high spatial and temporal resolution, operates over a wide temperature range and can be used to rapidly quantify local temperatures with high precision.
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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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Mello, Richard. "IDENTIFICATION OF DQ ALPHA POLYMORPHISM USING THE POLYMERASE CHAIN REACTION." VCU Scholars Compass, 1991. https://scholarscompass.vcu.edu/etd/5213.
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This was a study of detection systems for DQ alpha HLA polymorphism that could be exploited for the demonstration of simulated chimerism. Polymorphic segments of DO alpha DNA were amplified by the polymerase chain reaction (PCR). Simulated chimerism was represented by a mixture of minor and major component DNA. The goal was to detect minor component DNA in the presence of major component DNA utilizing various laboratory techniques. Techniques studied included probe strip typing with the AmpliType HLA-DO Alpha test kit, allele-specific amplification, polyacrylamide gel electrophoresis, restriction enzymes, and Southern transfer combined with a peroxidase detection system.
The AmpliType HLA-DO Alpha test kit had a detection sensitivity of at least 0.2%. This is much better than the 10% detection sensitivity in non-PCR techniques. When the 3.0 DC alpha type was mixed as the minor component with undiluted 1.1 DQ alpha type, the detection sensitivity for the 3.0 DC alpha type increased to a detection level of 0.1%.
The allele-specific primers were able to specifically amplify the minor component DNA in the presence of major component DNA. Major component DNA did not amplify and thus did not compete with the minor component DNA for Taq polymerase. The allele-specific primers provided an overall detection sensitivity of 0.2%.
Background interference prevented detection of minor component bands on both polyacrylamide gels stained with ethidium bromide and on Southern blots reacted with the peroxidase detection system.
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Tsang, Tsui-ying Stella. "Application of quantitative polymerase chain reaction in the diagnosis of thalassaemia /." View the Table of Contents & Abstract, 2006. http://sunzi.lib.hku.hk/hkuto/record/B36433895.
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Tsui, Wai-yan. "Determination of PTEN mutations in prostate cancer in Chinese." Hong Kong : University of Hong Kong, 2001. http://sunzi.lib.hku.hk/hkuto/record.jsp?B23736173.
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顔鴻儀 and Hung-yee Ngan. "Molecular diagnosis of penicilliosis marneffei." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2001. http://hub.hku.hk/bib/B31970035.
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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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Ngan, Hung-yee. "Molecular diagnosis of penicilliosis marneffei." Hong Kong : University of Hong Kong, 2001. http://sunzi.lib.hku.hk/hkuto/record.jsp?B23595978.
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McMullen, Kevin Patrick. "Inhibitory effects of food matrices on real-time reverse transcription polymerase chain reaction detection of foodborne viruses." [Tampa, Fla. : s.n.], 2003. http://purl.fcla.edu/fcla/etd/SFE0000100.
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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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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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Walker, Ken R. "Rapid detection of Listeria monocytogenes in salad by polymerase chain reaction." Auburn, Ala, 2005. http://repo.lib.auburn.edu/2005%20Summer/master's/WALKER_KEN_28.pdf.
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Phaneuf, Christopher. "Infrared laser-mediated polymerase chain reaction in a polymer microfluidic device." Diss., Georgia Institute of Technology, 2014. http://hdl.handle.net/1853/53068.
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The ability to rapidly, sensitively, and accurately detect the presence of a pathogen is a vital capability for first responders in the assessment and treatment of scenarios such as disease outbreak and bioterrorism. Nucleic acid tests such as the polymerase chain reaction (PCR) are supplanting traditional techniques due to the improved speed, specificity, sensitivity, and simplicity. Still, amplification by PCR is often the bottleneck when processing genetic samples. Conventional PCR machines are bulky, slow, and consume large reagent volumes and an affordable, compact, efficient, easy-to-use alternative has yet to emerge. In this work, a microfluidic PCR platform was developed consisting of a low-cost, multi-chamber polymer microchip and a laser-mediated thermocycler capable of independent thermal control of each reaction chamber. Innovations in polymer microchip modeling, fabrication, and characterization yielded a low-cost solution for sample handling. A simple optical system featuring an infrared laser diode and solenoid-driven optical shutter was combined with a microfluidic temperature measurement system utilizing embedded thermocouples to achieve rapid thermocycling capable of multiplexed temperature control. We validated the instrument with sensitive amplifications of multiple viral targets simultaneously. This technology is a breakthrough in practical microfluidic PCR instrumentation, providing the foundation for a paradigm shift in low-cost, high-throughput genetic diagnostics.
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Tully, Gillian. "DNA profiling for forensic identification : evaluation of polymerase chain reaction methods." Thesis, Cardiff University, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.264882.
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Diss, Timothy Charles. "The polymerase chain reaction in the characterisation and diagnosis of lymphomas." Thesis, University College London (University of London), 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.338653.
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Organji, Sameer R. A. "Detection of #Beta#-lactam resistant Streptococcus pneumoniae by polymerase chain reaction." Thesis, University of Bristol, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.297824.
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Sim, Steven Poh Chuen. "An integrated chip-based device for droplet-flow polymerase chain reaction." Thesis, Imperial College London, 2016. http://hdl.handle.net/10044/1/56111.
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The polymerase chain reaction (PCR) is an important in-vitro technique in molecular biology for amplifying trace quantities of deoxyribonucleic acid (DNA). PCR is carried out by mixing the DNA molecules to be amplified with primers, polymerase enzymes and deoxynucleotide triphosphates (dNTPs) in a suitable buffer solution. A conventional thermal-cycler is then used to cycle the PCR mixture between multiple temperatures for denaturation, annealing and extension. Bench-top thermal cyclers have large thermal masses and use large sample volumes, leading to overly long cycling times, excessive energy and material consumption, inhomogeneity in the reaction environment, and an inability to handle large numbers of small volume aliquots. Microfluidic technologies overcome many of the limitations of bench-top thermal cyclers, providing a more controlled approach to PCR. Droplet-flow is one of the most promising microfluidic methods for carrying out PCR. The droplet-flow approach uses small water-in-oil droplets for compartmentalisation of the PCR reaction mixture, with each droplet behaving like an individual reaction chamber. By flowing the droplets over different temperatures for denaturation, annealing and extension, rapid thermal cycling can be achieved, greatly reducing the reaction time relative to bench-top thermal cyclers. The use of an oil phase to encapsulate the aqueous PCR mixture as droplets also prevents unwanted surface interactions and flow dispersion that can adversely affect the PCR yield. Here we describe an integrated microfluidic device for carrying out droplet-flow PCR. Instead of using multiple temperature zones to thermally-cycle the flowing droplets, the device used an on-chip radial temperature gradient. The droplets passed through microchannels arranged in a spoke-like geometry, causing them to pass backwards and forwards along the radial temperature gradient and so undergo the repeated thermal cycling required for PCR. The device reported here builds on an earlier plastic microfluidic PCR device (by Schaerli et al.) in which the radial temperature gradient was generated using a bulky external heater and a thermoelectric cooler, together with heat sinks and fans. In the silicon- and glass-based device reported here, integrated heaters, temperature sensors and air gaps (for passive cooling) were used to generate the temperature gradient, leading to significant miniaturisation of the device. The dimensions of the complete device assembly were 6.0 cm x 5.0 cm x 2.0 cm compared to 25.0 cm x 25.0 cm x 25.0 cm for the device by Schaerli et al. Despite the small size of the device, the achievable temperature gradient on the chip was sizeable. For instance, when the central heater was set to 92.0 °C, the temperature at the periphery was ~60.0 °C, corresponding to a temperature difference of ~32.0 °C – easily sufficient for PCR applications. Using chemical modification, the hydrophilic walls of the microchannel were rendered hydrophobic. An on-chip T-junction or flow-focusing junction was subsequently used to merge the oil and aqueous streams to generate the PCR-containing water-in-oil droplets. A PCR recipe was optimised on a bench-top thermal cycle. With this recipe, droplet-flow PCR was conducted on the PCR device by flowing the generated droplets up and down the radial temperature gradient to induce thermal cycling. Gel electrophoresis analysis of the collected droplets from the device showed the presence of the PCR product, confirming the ability of the integrated device to conduct droplet-flow PCR. By varying the central temperature of the PCR device and the flow rate of the droplets, the yield of the PCR product could be tuned. By serially diluting the concentration of the DNA molecules, it was found that the PCR device was able to amplify concentrations as low as 0.01 pM to a level detectable by gel electrophoresis. When coupled to a laser-induced fluorescence detection system, the emission from the PCR mixture in the water-in-oil droplets could be successfully detected for each PCR cycle. The increase in the fluorescence over successive PCR cycles once again verified the feasibility of carrying out droplet-flow PCR on the integrated device.
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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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Holladay, Ervin Blair. "Viral gene detection in oral neoplasms using the polymerase chain reaction." The Ohio State University, 1991. http://rave.ohiolink.edu/etdc/view?acc_num=osu1345129906.
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Holladay, E. Blair. "Viral gene detection in oral neoplasms using the polymerase chain reaction /." The Ohio State University, 1991. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487694702781994.
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Liao, Yu-Hua. "Polymerase chain reaction based cloning of acetate kinase in Propionibacterium acidipropionici /." Connect to this title online, 2003. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=osu1072778140.
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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.
Full textAbstract:
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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Su, Ying-Tu, and 蘇英圖. "Application of MicroEmusification：Emulsion Polymerase Chain Reaction." Thesis, 2009. http://ndltd.ncl.edu.tw/handle/51940901593932821213.
Full textAbstract:
碩士國立臺灣海洋大學
機械與機電工程學系
97
In this thesis, soft lithography technique was used to produce a micro emulsion chip. This chip can handle tiny volume of DNA solution for polymerase chain reation (ePCR). The DNA solution is dispersed in the emulsified droplets in a continuous oil phase, and the droplets become reaction space for PCR. As a result, the consistency of the multiple types of DNA templates existing in the original reaction space can be improved by emulsification process. Also, it will reduce competitive sequence interference and improve DNA magnification. Differing from usual micro emulsification chips, this study focuses on the technical problems involved in dealing with tiny and expensive DNA solution in order to avoid non-uniform droplets in the transient of droplet generation. The main ideas are as follows: (1) Let water first go through transient process and produce water droplets; (2) Use water and bubble to place DNA solution in between, and pass it through emulsification hydrodynamic focusing channel; (3) Use pneumatic membrane valves control the stop and flow of water and DNAsolution. Characterization of the emulsification chips includes droplet sizes, uniformity of droplets and hot-resistance of water-in-oil droplets. The smallest droplet diameter is less than 10 μm and the coefficient of variation is about 2%. The resulting electrophoretograms of both single or multiple templates’ PCR demonstrate that ePCR can improve the precision of amplified DNA segments. It is suggested from the experiment results that ePCR was proven to be efficient and could be used in further genetic research Key word: ePCR, DNA template, transient process, tiny-volume micro-emulsification
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Gi, Kao Li, and 高麗姬. "ABO Genotyping for Mutiplex Polymerase Chain Reaction." Thesis, 1999. http://ndltd.ncl.edu.tw/handle/82466547226309854097.
Full textAbstract:
碩士中央警察大學
刑事警察研究所
87
Abstract ABO blood group is clinically the most important blood group system in transfusion medicine including fours common different phenotypes . It has been a valuable tool in transfusion medicine , physical anthropology , disputed parentage testing , human identification , and in forensic analysis . Historically , forensic and clinical labatories utilize serological techniques to identify ABO blood type . The testing of forensic samples for ABO types using serological techniques has several drawbacks . We has simplified the identification of ABO types by taking advantage of previouly reported ABO DNA sequence difference . The use of single strand conformation polymorphism（SSCP）can separate sequence polymorphism than differ by only one base . We has investigated in 260 Chinese donors by multiplexed Polymerase Chain Reaction . In this study the ABO alleles from exon 6 , producing a 212/213 bp fragment , and exon 7, which produces a fragment of 272 bp , were separated by non-denaturing polyacrylamide gels . The two exons were amplified in a single reaction that produces similar quantities of DNA for both exons . However , in 260 SSCP patterens we can clustered 9 into different distinct and discernible groups . A concordance rate of 98.5 ﹪（256/260 samples）observed between the actural genotype and the serologically-based predicted genotype . These results indicate that the assay provides a rapid , accurate , and simple method for ABO genotyping that serves as a useful supplement to standard serological ABO typing .
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Chen, Jhao-Rong, and 陳昭榮. "Micro Rayleigh-Bénard polymerase chain reaction system." Thesis, 2005. http://ndltd.ncl.edu.tw/handle/05888212591754965876.
Full textAbstract:
碩士國立清華大學
工程與系統科學系
93
Polymerase chain reaction (PCR) is a molecular biological method for the in vitro amplification of nucleic acid molecule. In this thesis, the research object was to design a micro PCR system which involved a Rayleigh-Bénard convection PCR chip, measurement circuits, and temperature control circuits. Rayleigh-Bénard convection PCR chip was easy to be fabricated, and the sample solution in it can transit its temperature immediately. Thus, the faster the speed of flow is, the higher heating and cooling rate is. Rayleigh-Bénard convection PCR chip can be divided into two parts. First part is a chip with micro heaters and micro temperature sensors. Second part is PDMS reaction chamber designed by Rayleigh-Bénard convection theory. Temperature control system is designed to keep temperature on the top and bottom chips by Pulse width modulation control. Analog temperature signal is precisely measured by circuits. Then the signal is processed by an 8051 single chip, and the output power of micro heaters is controlled to adjust the temperature of top and down plates. Finally, agarose gel electrophoresis is used to verify the practicability of Rayleigh-Bénard convection PCR system. By comparing with the PCR experiments done by the commercial PCR machine and our chips, our chips can plenty decrease the reaction time. And By comparing with the simulation and PCR experiments of different designed sizes, users can use setted parameters and CFD results to do optimal designs and then decrease the total reaction time in the future.
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Tao, Yo-Shen, and 刁宥升. "Design Of Polymerase Chain Reaction System Controller." Thesis, 2002. http://ndltd.ncl.edu.tw/handle/76827215729605694932.
Full textAbstract:
碩士國立暨南國際大學
電機工程學系
90
DNA analysis is a crucial process for biotechnology. Polymerase chain reaction (PCR) is an imperative step for performing DNA analysis. PCR can proliferate a small quantity of DNA sample to a large quantity enough for DNA analysis in a short time. Our goal is to develop a PCR system chip by integrating standard CMOS process and MEMS process. In this thesis, an 8-bit 4-level pipelined programmable micro-controller for temperature control during PCR process is designed by HDL description and implemented by using field programmable gate array (FPGA). The micro-controller includes modules of data path and control unit. Verilog hardware description language is used to model the modules. The individual module is functionally verified by Max Plus II of Altera and integrated into an 8-bit micro-controller. The micro-controller is realized by a FPGA chip, and then micro program for the temperature control of PCR is downloaded into the ROM in FPGA for system test. The micro-controller proposed in this thesis is very simple and easy to implement. It is suitable for general low speed applications. In order to easily integrate the micro-controller into other applications, we will try to parameterize its design variables for adjusting its hardware structure in future works.
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Gao, Li-Ji, and 高麗姬. "ABO Genotyping for Multiplex Polymerase Chain Reaction." Thesis, 1998. http://ndltd.ncl.edu.tw/handle/7433nn.
Full textAbstract:
碩士中央警察大學
刑事警察研究所
87
ABO blood group is clinically the most important blood group system in transfusion medicine including fours common different phenotypes . It has been a valuable tool in transfusion medicine , physical anthropology , disputed parentage testing , human identification , and in forensic analysis . Historically , forensic and clinical labatories utilize serological techniques to identify ABO blood type . The testing of forensic samples for ABO types using serological techniques has several drawbacks . We has simplified the identification of ABO types by taking advantage ofpreviouly reported ABO DNA sequence difference . The use of single strand conformation polymorphism ( SSCP ) can separate sequence polymorphism than differ by only one base . We has investigated in 260 Chinese donors by multiplexed Polymerase Chain Reaction . In this study the ABO alleles from exon 6 , producing a 212/213 bp fragment , and exon 7, which produces a fragment of 272 bp, were separated by non-denaturing polyacrylamide gels. The two exons were amplified in a single reaction that produces similar quantities of DNA for both exons . However , in 260 SSCP patterens we can clustered 9 into different distinct and discernible groups . A concordance rate of 98.5 % (256/260 samples) observed between the actural genotype and the serologically-based predicted genotype . These results indicate that the assay provides a rapid , accurate , and simple method for ABO genotyping that serves as a useful supplement to standard serological ABO typing.
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Lin, Min-Han. "PDMS (polydimethylsiloxane) Based Polymerase Chain Reaction Component Design." 2005. http://www.cetd.com.tw/ec/thesisdetail.aspx?etdun=U0001-2707200515463700.
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張澔任. "Detection of Chlamydophila abortus by polymerase chain reaction." Thesis, 2000. http://ndltd.ncl.edu.tw/handle/59334353234665564833.
Full textAbstract:
碩士國立中興大學
獸醫微生物學研究所
88
The purpose of this study was to develop a specific polymerase chain reaction for rapid detection of Chlamydophila abortus. We selected the published chlamydial omp A and 16S-23S rRNA gene sequences from GeneBank, and respectively aligned these sequences to find suitable regions for Chlamydophila abortus-specific primer design. The alignment of 16S-23S rRNA gene sequences revealed no suitable region for primer design, while the alignment of omp A gene sequences revealed many suitable regions for Chlamydophila abortus- specific primer design. We designed several primer pairs from these suitable regions of omp A gene by Primer_3, and finally we selected 358f-868r, 433f-834r, 835f-1064r and 402f-1064r four primer pairs for the examination of detection of Chlamydophila abortus by polymerase chain reaction. The result showed that all the four primer pairs were specific for Chlamydophila abortus, so that our polymerase chain reaction with these primer pairs could permit the rapid and specific detedtion of Chlamydophila abortus, and the 358f-868r primer pair was the most sensitive pair. By this study we have developed a specific polymerase chain reaction for rapid detection of Chlamydophila abortus.