Bibliographies thématiques / PCR real time quantitativa

Littérature scientifique sur le sujet « PCR real time quantitativa »

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Publié le 11 mars 2023

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Articles de revues sur le sujet "PCR real time quantitativa"

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Heid, C. A., J. Stevens, K. J. Livak et P. M. Williams. « Real time quantitative PCR. » Genome Research 6, no 10 (1 octobre 1996) : 986–94. http://dx.doi.org/10.1101/gr.6.10.986.

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Schmittgen, Thomas D. « Real-Time Quantitative PCR ». Methods 25, no 4 (décembre 2001) : 383–85. http://dx.doi.org/10.1006/meth.2001.1260.

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RAZA, ABIDA, et NAUREEN A KHATTAK. « REAL TIME PCR ; ». Professional Medical Journal 19, no 06 (3 novembre 2012) : 751–59. http://dx.doi.org/10.29309/tpmj/2012.19.06.2455.

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In recent years, real-time PCR has come forward as a robust and widely used molecular technique in clinical and biologicalsettings. Although it can detect very minute quantities of target nucleic acid, but quantification of specific nucleic acids is not an easy task.Accurate and precise quantification is hampered by a number of factors that may include assay development and validation, fluorophoresselection, handling during sample preparation, storage, reaction procedures, and batch analysis conditions. Even minor variations aresignificantly magnified by the exponential nature of this technique. Current review gives an insight of the advantages, limitations, assaychemistries, quantitation parameters, and quality control issues related to this technology. Moreover it will also highlight the utilization of Realtime PCR in clinical oncology, virology, microbiology, and gene expression studies.
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Choi, Yeon-Jae, Sun-Ho Kim, Min-Jeong Gu, Han-Na Choe, Dong-Un Kim, Sang-Bum Cho, Su-Ki Kim, Che-Ok Jeon, Gui-Seok Bae et Sang-Seok Lee. « Quantitative Real-time PCR using Lactobacilli as Livestock Probiotics ». Journal of Life Science 20, no 12 (30 décembre 2010) : 1896–901. http://dx.doi.org/10.5352/jls.2010.20.12.1896.

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Bell, Andrew S., et Lisa C. Ranford-Cartwright. « Real-time quantitative PCR in parasitology ». Trends in Parasitology 18, no 8 (août 2002) : 338–42. http://dx.doi.org/10.1016/s1471-4922(02)02331-0.

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Wong, Marisa L., et Juan F. Medrano. « Real-time PCR for mRNA quantitation ». BioTechniques 39, no 1 (juillet 2005) : 75–85. http://dx.doi.org/10.2144/05391rv01.

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Bustin, S. A., V. Benes, T. Nolan et M. W. Pfaffl. « Quantitative real-time RT-PCR – a perspective ». Journal of Molecular Endocrinology 34, no 3 (juin 2005) : 597–601. http://dx.doi.org/10.1677/jme.1.01755.

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The real-time reverse transcription polymerase chain reaction (RT-PCR) uses fluorescent reporter molecules to monitor the production of amplification products during each cycle of the PCR reaction. This combines the nucleic acid amplification and detection steps into one homogeneous assay and obviates the need for gel electrophoresis to detect amplification products. Use of appropriate chemistries and data analysis eliminates the need for Southern blotting or DNA sequencing for amplicon identification. Its simplicity, specificity and sensitivity, together with its potential for high throughput and the ongoing introduction of new chemistries, more reliable instrumentation and improved protocols, has made real-time RT-PCR the benchmark technology for the detection and/or comparison of RNA levels.
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Sochivko, D. G., A. A. Fedorov, D. A. Varlamov, V. E. Kurochkin et R. V. Petrov. « Accuracy of quantitative real-time PCR analysis ». Doklady Biochemistry and Biophysics 449, no 1 (mars 2013) : 105–8. http://dx.doi.org/10.1134/s1607672913020154.

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Arya, Manit, Iqbal S. Shergill, Magali Williamson, Lyndon Gommersall, Neehar Arya et Hitendra RH Patel. « Basic principles of real-time quantitative PCR ». Expert Review of Molecular Diagnostics 5, no 2 (mars 2005) : 209–19. http://dx.doi.org/10.1586/14737159.5.2.209.

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Boulay, J. L., J. Reuter, R. Ritschard, L. Terracciano, R. Herrmann et C. Rochlitz. « Gene Dosage by Quantitative Real-Time PCR ». BioTechniques 27, no 2 (août 1999) : 228–32. http://dx.doi.org/10.2144/99272bm03.

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Thèses sur le sujet "PCR real time quantitativa"

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Andalo, Alice. « Analisi quantitativa dell'espressione genica mediante real-time rt-pcr ». Bachelor's thesis, Alma Mater Studiorum - Università di Bologna, 2015. http://amslaurea.unibo.it/8450/.

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Il primo capitolo di questo lavoro di tesi introduce i concetti di biologia necessari per comprendere il fenomeno dell’espressione genica. Il secondo capitolo descrive i metodi e le tecniche di laboratorio utilizzate per ottenere il cDNA, il materiale genetico che verrà amplificato nella real-time PCR. Nel terzo capitolo si descrive la tecnica di real-time PCR, partendo da una descrizione della PCR convenzionale fino a delineare le caratteristiche della sua evoluzione in real-time PCR. Si prosegue con la spiegazione del principio fisico alla base della tecnica e delle molecole necessarie (fluorofori e sonde) per realizzarla; infine si descrive l’hardware e il software dello strumento. Il quarto capitolo presenta le tecniche di analisi del segnale che utilizzano metodi di quantificazione assoluta o relativa. Infine nel quinto capitolo è presentato un caso di studio, cioè un’analisi di espressione genica con real-time PCR condotta durante l’esperienza di tirocinio presso il laboratorio ICM. e delle molecole necessarie (fluorofori e sonde) per realizzarla; infine si descrive l’hardware e il software dello strumento. Il quarto capitolo presenta le tecniche di analisi del segnale che utilizzano metodi di quantificazione assoluta o relativa. Infine nel quinto capitolo è presentato un caso di studio, cioè un’analisi di espressione genica con real-time PCR condotta durante l’esperienza di tirocinio presso il laboratorio ICM.
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Chagas, Júnior Adenizar Delgado das. « Avanços no conhecimento da imunopatogênese da leptospirose e a aplicação do método do imprint como ferramenta qualitativa e quantitativa de leptospiras ». Centro de Pesquisas Gonçalo Moniz, 2014. https://www.arca.fiocruz.br/handle/icict/7628.

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Submitted by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2014-05-20T17:14:19Z No. of bitstreams: 1 Adenizar Delgado das Chagas Jr. Avanços... 2014..pdf: 11153892 bytes, checksum: 48154dea65a7a4a32a54408e694c5d30 (MD5)
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Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil
A leptospirose é uma zoonose causada por espiroquetas patogênicas pertencentes ao gênero Leptospira. O modelo da doença em camundongos tem vantagens devido à ampla gama de ferramentas genéticas e imunológicas disponíveis para pesquisas básicas. A maior limitação na conduta clínica e na pesquisa experimental da leptospirose é o fraco desempenho dos métodos disponíveis para detecção direta e para quantificação de leptospiras. Foi incluído nesta tese um conjunto de três manuscritos que visam investigar o desfecho da infecção pela cepa virulenta de Leptospira interrogans nas linhagens de camundongos selvagens (A, CBA, BALB/c e C57BL/6), em camundongos óxido nítrico sintase induzível (iNOS) Knockout (KO), camundongos gene ativador de recombinação 1 (RAG1) KO , camundongos CB17 com imunodeficiência combinada grave (SCID), e os seus respectivos controles selvagens C57BL/6 e BALB/c. Investigar a confiabilidade do método de quantificação do imprint (IM), comparando os resultados obtidos com esta técnica aos obtidos com a utilização do PCR em tempo real (qPCR) para detectar e quantificar leptospiras em amostras de rim de ratos e hamsters experimentalmente infectados. Como esperado, nenhuma das linhagens de camundongos selvagens foram suscetíveis à leptospirose letal. A linhagem A e C57BL/6 exibiram altas cargas de Leptospira nas amostras de rim e as linhagens CBA e C57BL/6 desenvolveram lesões inflamatórias graves, enquanto a linhagem BALB/c provou ser a mais resistente apresentando leptospirose subclínica. Os camundongos iNOS KO e selvagem sobreviveram sem sintomas clínicos de leptospirose. A frequência e gravidade das nefrites foram significantemente menores nos camundongos iNOS KO. Todos os animais RAG1 KO e SCID morreram de leptospirose aguda, enquanto que todos os camundongos selvagens sobreviveram. A hemorragia pulmonar foi observada em 57 e 94% dos camundongos RAG 1 KO e em 83 e 100% dos camundongos SCID, usando doses de inóculos de 107 e 106 leptospiras, respectivamente. Não houve evidências de hemorragia pulmonar nos controles selvagens. Nos modelos de infecção agudo e crônico, houve correlação positiva estatisticamente significante (P < 0,05) na quantificação de leptospiras pelos métodos do qPCR e do IM. Como conclusão geral, a linhagem de camundongos A pode ser a linhagem de escolha em estudos na qual se pretende recuperar um grande número de leptospiras de rins colonizados. As linhagens CBA e C57BL/6 desenvolveram, com maior frequência, lesões inflamatórias e podem ser as mais adequadas para estudos de leptospirose associados com nefrite intersticial. A linhagem BALB/c é a mais indicada para estudar mecanismos que envolvam a imunidade inata e/ou a rápida resposta imune adaptativa. A ausência do gene do iNOS no modelo murino resultou em uma diminuição significativa da suscetibilidade para o desenvolvimento da nefrite intersticial. Além disso, a ausência de linfócitos B e T funcionais não impediu a ocorrência de hemorragia pulmonar. Estes dados fornecem fortes evidências de que a hemorragia pulmonar na leptospirose não está relacionada apenas a mecanismos autoimunes. Para a detecção e quantificação de leptospiras o método do imprint foi equivalente ao qPCR.
Leptospirosis is a zoonosis caused by pathogenic spirochaetes belonging to the genus Leptospira. The mouse disease model is advantagous due to the broad array of immunological and genetic tools available for basic research. A major limitation in the clinical management and experimental research of leptospirosis is the poor performance of the available methods in the direct detection and quantification of leptospires. This thesis includes three manuscripts that investigate the outcome of infection by a virulent strain of Leptospira interrogans in wildtype mice strains: A, CBA, BALB/c and C57BL/6; in iNOS knockout (KO) mice, recombination activating gene 1 (RAG1) KO mice and CB17 severe combined immunodeficiency (SCID) mice. To investigate whether the imprint method (IM) of quantification was reliable we compared it with against real time PCR (qPCR) for the detection and quantification of leptospires in kidney samples from rats and hamsters. As expected, none of the wildtype mice were susceptible to lethal leptospirosis. The A and C57BL/6 strains exhibited high leptospiral loads in the kidney samples and the CBA and C57BL/6 strains developed severe inflammatory lesions, whilst the BALB/c strain proved to be the most resistant to subclinical leptospirosis. The iNOS KO mice survived with no clinical symptoms of leptospirosis. The frequency and severity of nephritis was significantly lower in the iNOS KO mice. All of the RAG 1 KO and SCID animals died of acute leptospirosis, whereas all of the wildtype mice survived. Pulmonary haemorrhage was observed in 57 and 94% of Rag1 KO mice and in 83 and 100% of SCID mice, using inoculum doses of 107 and 106 leptospires, respectively. There was no evidence of pulmonary haemorrhage in the wildtype controls. In both the acute and chronic infection models, the correlation between quantification by qPCR and the IM was found to be positive and statistically significant (P < 0.05). In conclusion, the mouse A strain would be the strain of choice in studies requiring the recovery of a large number of leptospires from colonized kidneys. The CBA and C57BL/6 strains developed more inflammatory lesions and would be the most suitable for studies of leptospirosis associated with interstitial nephritis. The BALB/c strain appeared to be the most suitable for studying mechanisms involving innate immunity and/or rapid adaptive immune response. The absence of the iNOS gene in the murine model resulted in a significant decrease in susceptibility to the development of interstitial nephritis. Furthermore, the absence of functional B and T lymphocytes did not prevent the occurrence of pulmonary hemorrhage. These data provide strong evidence that pulmonary hemorrhage in leptospirosis is not only related to autoimmune mechanisms. For the detection and quantification of leptospires the imprint method was quivalent to that of qPCR. Keywords:
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Zhang, Yan. « Frequent RASSF1A gene promoter hypermethylation in breast cancer ». [S.l. : s.n.], 2008. http://nbn-resolving.de/urn:nbn:de:bsz:289-vts-63611.

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Silva, Thaís Galhardo Egreja Ribeiro da. « Raquitismo da soqueira de cana-de-açúcar : transmissão do patógeno via sementes e avaliação quantitativa de seu controle através da termoterapia de toletes ». Universidade de São Paulo, 2013. http://www.teses.usp.br/teses/disponiveis/11/11135/tde-17122013-112133/.

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Devido à sua múltipla utilidade, a cana-de-açúcar (Saccharum spp.) é amplamente cultivada em mais de 127 países tropicais e subtropicais. No Brasil, a cultura apresentou expansão em área plantada nos últimos anos em virtude da crescente demanda por biocombustíveis. Porém, a produção de mudas sadias tende a ser um fator limitante a este aumento de demanda, já que diversas doenças são transmitidas por toletes contaminados. Dentre estas, destaca-se o raquitismo da soqueira (RSD), causado pela bactéria fastidiosa Leifsonia xyli subsp. xyli (Lxx) (KAO e DAMANN, 1980). De acordo com a literatura, a transmissão de Lxx se dá através de toletes contaminados e/ou instrumentos de corte, não havendo relatos sobre sua transmissão por sementes. Desta forma, seu controle se baseia no princípio da exclusão, através do uso de toletes tratados termicamente como fonte de material propagativo. Porém, não se conhece o efeito do tratamento no título de Lxx, uma vez que os relatos de eficiência da termoterapia baseiam-se em avaliações qualitativas. Em face destas informações, este estudo objetivou avaliar a transmissão de Leifsonia xyli subsp. xyli por sementes e também quantificar o efeito da termoterapia no título de Lxx em duas variedades de cana-de-açúcar através da PCR quantitativa em tempo real. No ensaio de transmissão via sementes, a bactéria foi detectada em altas incidências (>87%) em plantas de 90 dias oriundas de sementes coletadas de plantas infectadas e em quantidades estimadas superiores a 51 células por 100ng de DNA total de sementes. Além disso, aos 270 dias após o plantio, a bactéria foi isolada de 6 plantas. A identidade das colônias foi comprovada através de PCR convencional utilizando primers específicos para Lxx. Dois ensaios foram conduzidos para quantificar o efeito da termoterapia na população bacteriana. O título de Lxx foi determinado em folhas de plantas de duas variedades de cana-deaçúcar oriundas de toletes tratados termicamente ou não aos 90 dias após o plantio. O título inicial presente nos colmos utilizados como fonte de toletes interferiu na eficiência do tratamento, que reduziu a população bacteriana somente no ensaio onde os colmos apresentaram elevado título bacteriano inicial. Além disso, 70 e 55% das plantas oriundas de toletes tratados termicamente no primeiro e no segundo ensaio, respectivamente, apresentavam células de Lxx, porém em menores quantidades quando comparadas a plantas oriundas de toletes não tratados. Com isso, conclui-se que Lxx é transmitida em elevada frequência via sementes e a termoterapia, embora não tenha efeito erradicante pronunciado, é capaz de reduzir o título bacteriano no tecido vegetal.
Due to its multiple uses, sugarcane (Saccharum spp.) is widely cultivated in more than 127 tropical and subtropical countries. In Brazil, sugarcane cropping has increased in recent years because of the growing demand for biofuels. However, the production of healthy seedlings tends to be a limiting factor to this increase, since contaminated setts transmit many diseases. Among these, the Ratoon Stunting Disease (RSD) is a major disease caused by the fastidious bacterium Leifsonia xyli subsp. xyli (Lxx) (KAO and DAMANN, 1980). According to the literature, the transmission of Lxx occurs through contaminated setts and/or cutting tools, but there are no reports of its transmission by seeds. Therefore, its control is based on the principle of exclusion using heat-treated setts as source of propagative material. However, the quantitative effect of the treatment on the bacterial titer is not known, since reports of thermotherapy efficiency are based on qualitative assessments. This study aimed to evaluate the transmission of Leifsonia xyli subsp. xyli by seeds and to quantify the effect of thermotherapy on Lxx in two sugarcane varieties using real-time quantitative PCR. In the test of disease transmission by seeds, the bacterium was detected in high incidences (>87%) in 90-day old plants generated from seeds collected from infected plants in quantities exceeding 51 cells per 100ng of total DNA of seeds. In addition, at 270 days after planting, the bacterium was isolated from six plants. The colony identity was confirmed by conventional PCR using Lxx-specific primers. Two trials were conducted to quantify thermotherapy effect on bacterial population. Lxx titers were determined 90 days after planting in plant leaves of two sugarcane varieties originated from setts subjected or not to heat-treatment. The initial bacterial titer in the stalks affected the treatment efficiency, which significantly reduced Lxx titers only in the trial where the stalks presented high initial bacterial levels. Moreover, 70 and 55% of the plants originated from heat-treated setts in the first and second tests, respectively, were infected with Lxx, cells; however, in smaller amounts compared to plants from untreated setts. Thus, it is concluded that Lxx is transmitted in high frequency by seeds and that the thermo treatment, although not efficient as an eradicating treatment, reduces the bacterial population in the plant tissue.
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Souza, William Marciel de. « Estudo evolutivo dos hantavírus e desenvolvimento de uma RT-PCR quantitativa em tempo real para detecção do vírus Araraquara ». Universidade de São Paulo, 2013. http://www.teses.usp.br/teses/disponiveis/17/17147/tde-10042013-123852/.

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O gênero Hantavírus está incluído na família Bunyaviridae que são vírus emergentes associados a roedores que podem infectar o homem causando graves doenças. Nas Américas, os Hantavírus causam uma síndrome pulmonar e cardiovascular (SPCVH) com alta letalidade. Cerca de 1600 casos de SPCVH já foram notificados no Brasil causando mais de 600 óbitos. Sete espécies de Hantavírus são conhecidas no Brasil incluindo o vírus Araraquara que circula nas regiões de cerrado do país associado ao roedor Necromys lasiurus. Para o desenvolvimento de uma RT-PCR em tempo real para detecção e quantificação de Hantavírus, mostramos as etapas para o desenvolvimento de uma one-step RT-PCR em tempo real SYBR Green I para Hantavírus Araraquara que se mostrou específica para o gênero e capaz de detectar até 10 cópias por mL de RNA viral na amostra. Além disso, realizamos um estudo filogenético utilizando algoritmos bayesianos, com 190 sequências completas do gene da nucleoproteína, oriundas de 30 países durante um período de 25 anos (1985-2010) que encontravam-se disponíveis no GenBank (NCBI). Baseando-se em uma taxa média de 6.8 x 10-4 (2.5 x 10-4 - 1 x 10-3) substituições nucleotídicas por sítio/ano, foi possível inferir que os Hantavírus teriam aproximadamente 1917 anos. O processo de dispersão dos Hantavírus pelo mundo teria ocorrido há aproximadamente 500 anos, e a introdução destes vírus nas Américas teria ocorrido há 549 anos (95% HPD 1555-341 anos), via América Central ou México, originando os Hantavírus adaptados aos roedores da subfamília Neotominae, e pelo Brasil surgindo há 406 anos (95% HPD 1150-250 anos) os Hantavírus associados a roedores da subfamília Sigmodontinae, e posteriormente dispersaram para todo o continente sul-americano. O trabalho contribui de forma relevante para o diagnóstico das infecções por Hantavírus com a one-step RT-PCR em tempo real SYBR Green I e também, contribui para o entendimento da filogenia e história destes vírus, oferecendo subsídios ao entendimento sobre como teria ocorrido o espalhamento dos Hantavírus pelo mundo.
The genus Hantavirus is included in the family Bunyaviridae are viruses emerging carried by rodents, which can infect humans causing serious illness. In the Americas, the Hantavirus causing a pulmonary syndrome (HPS) with high lethality. About 1,600 cases of HPS have been reported in Brazil, cause over 1600 deaths. Seven species of Hantavirus are known in Brazil, including Araraquara virus circulating in Cerrado regions (or Savannah regions) of the related in rodents Necromys lasiurus. The development of a real-time RT-PCR for detection and quantitation of Araraquara virus, here we show the steps for developing a one-step SYBR Green real-time RT-PCR for virus Araraquara which proved to be specific for the genus and capable of detecting up to 10 copies of viral RNA per ml in the sample. Furthemore, we performed a phylogenetic analysis using Bayesian algorithms, with 190 complete sequences of the nucleoprotein gene, originating from 30 countries over a 25 year period (1985-2010) that were available in GenBank (NCBI). Based on an average rate of 6.8 x 10-4 (2.5 x 10-4 - 1 x 10-3) nucleotide substitutions per site/year, it was possible to infer that the Hantavirus would be about 1917 years old. The Hantavirus spreading in the world have occurred for nearly 500 years, and the introduction of these viruses have occurred in the Americas 549 years ago (95 years% HPD 1555-341) bye Central America or Mexico, causing the Hantavirus adapted to rodents subfamily Neotominae, and Brazil emerged 406 years ago (95% HPD 1150-250 years) the Hantavirus associated with rodents subfamily Sigmodontinae, and subsequently disseminated to South America. The work contributes significantly to the diagnosis of Hantavirus infections with one-step SYBR Green real-time RT-PCR and also contributes to an understanding of the phylogeny and evolutionary history of these viruses, offering subsidies have occurred understanding of how the Hantavirus spread of the worldwide.
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ALBUQUERQUE, Yvana Maria Maia de. « Reação em cadeia da polimerase em tempo real quantitativa (QPCR) para diagnóstico da tuberculos pulmonar em escarro de pacientes com HIV/aids ». Universidade Federal de Pernambuco, 2012. https://repositorio.ufpe.br/handle/123456789/18503.

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O diagnóstico da tuberculose apresenta dificuldades em pacientes soropositivos para o vírus da imunodeficiência humana (HIV). Os doentes coinfectados HIV/M.tuberculosis(MTB)nos estágios mais avançados de imunocomprometimento apresentam manifestações clínicas atípicas e o exame direto, rotineiramente utilizado, tem baixa sensibilidade. A cultura apesar de ter maior sensibilidade fornece resultados tardios. Evidencia-se nesta população a necessidade de testes diagnósticos mais eficientes. A tese será apresentada no formato de dois artigos científicos. No primeiro, estudou-se a utilidade da Reação em Cadeia da Polimerase em tempo real quantitativa (qPCR) para diagnóstico da tuberculose pulmonar em escarro de pacientes com HIV/aids. No segundo, descreveu-se as alterações radiográficas do tórax de pacientes com tuberculose pulmonar confirmado por cultura de escarro. Foram incluídos no estudo 140 pacientes com HIV/aids e suspeita clínica de tuberculose pulmonar, atendidos no período de agostode 2009 a janeiro de 2012, em dois hospitais de referência para atendimento de pacientes infectados pelo HIV em Recife-PE. Coletou-se uma amostra de escarro de cada paciente, e caso não houvesse escarro suficiente, realizou-se nebulização com solução salina para indução do escarro. O padrão ouro do estudo foi à cultura realizada em meios Löwenstein-Jensen e 7H9. A cultura e a qPCR para tuberculose foram realizadas em laboratório privado situado no Recife. Dos 140 pacientes em 47 (33,6%), diagnosticou-se tuberculose pulmonar pelo padrão ouro. A sensibilidade, especificidade e acurácia da qPCR foram respectivamente 87,2%, 98,9% e 95%. Foram realizados exames radiográficos do tórax em 42 pacientes coinfectados com cultura de escarro positiva, que foram avaliados por dois radiologistas experientes. A alteração radiológica isolada mais frequente observada foi a consolidação parenquimatosa, que acometeu seis (14,3%) dos pacientes, seguida pelo infiltrado intersticial e micronodular difuso, além da associação infiltrado e consolidação. Concluiu-se que a qPCR realizada no escarro de pacientes coinfectados HIV/MTB apresentou boa sensibilidade, especificidade e acurácia, sendo útil no diagnóstico de tuberculose pulmonar nesses pacientes. Com relação aos achados radiográficos de tórax, estes demonstraram ser de pouco auxílio no diagnóstico da tuberculose pulmonar nos coinfectados, exceto quando o padrão cavidade e infiltrado micronodular difuso estão presentes.
Tuberculosis’ diagnosis is difficult in HIV soropositivepatients. The patients co-infected HIV/M.tuberculosis(MTB) in severe immune deficiency stage present atypical clinical manifestation and direct sputum smear, usually used, shows low sensitivity.The culture, despite better sensitivity, obtains later results. The thesis will be presented in form of two articles. In the first, it has studied the utility of quantitative real time PCR for tuberculosis’ diagnosis among AIDS patients’ sputum smear. In the second, it has been described the major thoracic radiographic alterations among AIDS patients’ and pulmonary tuberculosis confirmed by sputum culture. A total of 140 HIV/AIDS patients were included in the study, with clinical suspicion of pulmonary tuberculosis, from August 2009 to January 2012, were attended at two referral hospitals for HIV/AIDS in Recife-PE. A sputum sample was collected from each patient, and if they were unable to produce spontaneous sputum, they were saline nebulized, for sputum induction. The culture, in Löwenstein-Jensen solid medium and 7H9, was the gold standard. The culture and qPCR for tuberculosis have been done in a private laboratory in Recife-PE. From all the 140 studied patients, 47 (33,6%) pulmonary tuberculosis was diagnosis by gold standard. The sensitivity, specificity and accuracy were 87,7%, 98,9% and 95% respectively. There were evaluated chest X-rays from 42 co-infected HIV/MTB patients with positive culture by two experienced radiologists, the most common radiological alteration was parenchymal consolidation, encountered in six (14,3%) patients, followed by interstitial infiltrate, difuse micronodular (military) pattern an association between interstitial infiltrate and parenchymal consolidation. It was concluded that qPCR, has given a good sensitivity, specificity and accuracy and it can be recommender for use in the management of HIV/AIDS patients. However, thoracic radiographic findings were not specific and thorax RX is not sufficient initself to establish a pulmonary tuberculosis diagnosis in co-infected HIV/MTB patients, except when cavity and micronodular pattern are presented.
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Steyn, HC, A. Pretorius et CME McCrindle. « A quantitative real-time PCR assay for Ehrlichia ruminantium using pCS20 ». Elsevier, 2008. http://encore.tut.ac.za/iii/cpro/DigitalItemViewPage.external?sp=1000379.

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Heartwater is a tick borne disease that affects ruminants and wild animals in Africa south of the Sahara. It is caused by Ehrlichia ruminantium and transmitted by the tick Amblyomma hebraeum. The protocols currently used to detect heartwater take several days to complete. Here, we describe the development of a pCS20 quantitative real-time PCRTaqMan probe assay to detect E. ruminantium in livestock blood and ticks from the field. The assay is based on the conserved pCS20 gene region of E. ruminantium that contains two overlapping genes, rnc and ctaG [Collins, N.E., Liebenberg, J., De Villiers, E.P., Brayton, K.A., Louw, E., Pretorius, A., Faber, F.E., Van Heerden, H., Josemans, A., Van Kleef, M., Steyn, H.C., Van Strijp, M.F., Zweygarth, E., Jongejan, F., Maillard, J.C., Berthier, D., Botha, M., Joubert, F., Corton, C.H., Thomson, N.R., Allsopp, M.T., Allsopp, B.A., 2005. The genome of the heartwater agent Ehrlichia ruminantium contains multiple tandem repeats of actively variable copy number. PNAS 102, 838–843]. The pCS20 quantitative real-time PCRTaqMan probe was compared to the currently used pCS20 PCR and PCR/32P-probe test with regards to sensitivity, specificity and the ability to detect DNA in field samples and in blood from experimentally infected sheep. This investigation showed that the pCS20 quantitative real-time PCRTaqMan probe was the most sensitive assay detecting seven copies of DNA/ml of cell culture. All three assays, however, cross react with Ehrlichia canis and Ehrlichia chaffeensis. The pCS20 real-time PCR detected significantly more positive field samples. Both the PCR and pCS20 real-time PCR could only detect E. ruminantium parasites in the blood of experimentally infected sheep during the febrile reaction. The PCR/32P-probe assay, however, detected the parasite DNA 1 day before and during the febrile reaction. Thus, because this new quantitative pCS20 real-time PCRTaqMan probe assay was the most sensitive and can be performed within 2 h it is an effective assay for epidemiological surveillance and monitoring of infected animals.
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Tichopád, Aleš. « Quantitative real-time RT-PCR based transcriptomics improvement of evaluation methods / ». [S.l.] : [s.n.], 2004. http://deposit.ddb.de/cgi-bin/dokserv?idn=972765069.

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Leopold, Luciana Eleanor Dittmer Dirk Peter. « Development of real-time PCR assays for the quantitative detection of herpesviruses ». Chapel Hill, N.C. : University of North Carolina at Chapel Hill, 2008. http://dc.lib.unc.edu/u?/etd,1487.

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Thesis (M.S.)--University of North Carolina at Chapel Hill, 2008.
Title from electronic title page (viewed Sep. 16, 2008). "... in partial fulfillment of the requirements for the degree of Master of Science in the Curriculum of Genetics and Molecular Biology." Discipline: Genetics and Molecular Biology; Department/School: Medicine.
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Rosa, Stefanie Ulrike. « "Real-Time-PCR-Untersuchungen zur Persistenz von infektiösen Toxoplasma-gondii-Dauerstadien in Rohwurst-Erzeugnissen" ». Giessen VVB Laufersweiler, 2009. http://d-nb.info/996004408/04.

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Livres sur le sujet "PCR real time quantitativa"

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Biassoni, Roberto, et Alessandro Raso, dir. Quantitative Real-Time PCR. New York, NY : Springer New York, 2014. http://dx.doi.org/10.1007/978-1-4939-0733-5.

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Biassoni, Roberto, et Alessandro Raso, dir. Quantitative Real-Time PCR. New York, NY : Springer New York, 2020. http://dx.doi.org/10.1007/978-1-4939-9833-3.

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Quantitative real-time PCR in applied microbiology. Norfolk, UK : Caister Academic Press, 2012.

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Quantitative real-time PCR : Methods and protocols. New York : Humana Press, 2014.

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Tevfik, Dorak M., dir. Real-time PCR. New York : Taylor & Francis, 2006.

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Tevfik, Dorak M., dir. Real-time PCR. New York : Taylor & Francis, 2006.

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Meuer, Stefan, Carl Wittwer et Kan-Ichi Nakagawara, dir. Rapid Cycle Real-Time PCR. Berlin, Heidelberg : Springer Berlin Heidelberg, 2001. http://dx.doi.org/10.1007/978-3-642-59524-0.

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Julie, Logan, Edwards Kirstin et Saunders Nick, dir. Real-time PCR : Current technology and applications. Norfolk, UK : Caister Academic Press, 2009.

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Dietmaier, Wolfgang, Carl Wittwer et Natarajan Sivasubramanian, dir. Rapid Cycle Real-Time PCR — Methods and Applications. Berlin, Heidelberg : Springer Berlin Heidelberg, 2002. http://dx.doi.org/10.1007/978-3-642-59397-0.

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Wittwer, Carl, Meinhard Hahn et Karen Kaul, dir. Rapid Cycle Real-Time PCR — Methods and Applications. Berlin, Heidelberg : Springer Berlin Heidelberg, 2004. http://dx.doi.org/10.1007/978-3-642-18840-4.

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Chapitres de livres sur le sujet "PCR real time quantitativa"

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Sluijter, J. P. G., G. Pasterkamp et D. P. V. de Kleijn. « Quantitative Real-Time PCR ». Dans Cardiovascular Research, 75–83. Boston, MA : Springer US, 2006. http://dx.doi.org/10.1007/0-387-23329-6_4.

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Broll, Hermann. « Quantitative Real-Time PCR ». Dans Molecular Biological and Immunological Techniques and Applications for Food Chemists, 59–83. Hoboken, NJ, USA : John Wiley & Sons, Inc., 2010. http://dx.doi.org/10.1002/9780470637685.ch3.

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Dotti, Isabella, Ermanno Nardon, Danae Pracella et Serena Bonin. « Quantitative Real-Time RT-PCR ». Dans Guidelines for Molecular Analysis in Archive Tissues, 121–32. Berlin, Heidelberg : Springer Berlin Heidelberg, 2011. http://dx.doi.org/10.1007/978-3-642-17890-0_25.

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Martinez, Jeanelle M., et Nigel J. Walker. « Real-Time and Quantitative PCR ». Dans Toxicogenomics, 147–63. Hoboken, NJ, USA : John Wiley & Sons, Inc., 2005. http://dx.doi.org/10.1002/0471669040.ch7.

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Dötsch, Jörg, Ellen Schoof et Wolfgang Rascher. « Quantitative TaqMan Real-Time PCR ». Dans Medical Biomethods Handbook, 305–13. Totowa, NJ : Humana Press, 2005. http://dx.doi.org/10.1385/1-59259-870-6:305.

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Pal, Aruna. « Real-Time or Quantitative PCR ». Dans Springer Protocols Handbooks, 181–209. New York, NY : Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1818-9_9.

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Singh, Charanjeet, et Sinchita Roy-Chowdhuri. « Quantitative Real-Time PCR : Recent Advances ». Dans Clinical Applications of PCR, 161–76. New York, NY : Springer New York, 2016. http://dx.doi.org/10.1007/978-1-4939-3360-0_15.

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Honda, Junichi, et Kotaro Oizumi. « Quantitative Analysis of CMV in Infected Mice on the LightCycler System ». Dans Rapid Cycle Real-Time PCR, 349–57. Berlin, Heidelberg : Springer Berlin Heidelberg, 2001. http://dx.doi.org/10.1007/978-3-642-59524-0_37.

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Krüger, Petra, Albrecht Wiedenmann, Despina Tougianidou et Konrad Botzenhart. « Quantitative Detection of Cryptosporidium parvum after In Vitro Excystation by LightCycler PCR ». Dans Rapid Cycle Real-Time PCR, 341–48. Berlin, Heidelberg : Springer Berlin Heidelberg, 2001. http://dx.doi.org/10.1007/978-3-642-59524-0_36.

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Caplin, Brian Erich. « Quantification of Human Papilloma Virus Type 16 Using Quantitative Competitive PCR on the LightCycler ». Dans Rapid Cycle Real-Time PCR, 57–64. Berlin, Heidelberg : Springer Berlin Heidelberg, 2001. http://dx.doi.org/10.1007/978-3-642-59524-0_6.

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Actes de conférences sur le sujet "PCR real time quantitativa"

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Woudenberg, Timothy M., et J. Stevens. « Quantitative PCR by real-time detection ». Dans Photonics West '96, sous la direction de Gerald E. Cohn, Steven A. Soper et C. H. Winston Chen. SPIE, 1996. http://dx.doi.org/10.1117/12.237619.

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« RqPCRAnalysis : Analysis of Quantitative Real-time PCR Data ». Dans International Conference on Bioinformatics Models, Methods and Algorithms. SciTePress - Science and and Technology Publications, 2013. http://dx.doi.org/10.5220/0004312002020211.

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Ma, Yutong, Liang Zeng et Jianhuan Zhang. « A fluorescence detection optical system for real-time quantitative PCR ». Dans Optical Design and Testing X, sous la direction de Rengmao Wu, Osamu Matoba, Yongtian Wang et Tina E. Kidger. SPIE, 2020. http://dx.doi.org/10.1117/12.2574901.

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« Quantitative real-time PCR as a supplementary tool for molecular cytogenetics ». Dans Plant Genetics, Genomics, Bioinformatics, and Biotechnology. Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, 2019. http://dx.doi.org/10.18699/plantgen2019-044.

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Sayers, Michael B., et Tara M. Dalton. « A Real-Time Continuous Flow Polymerase Chain Reactor for DNA Expression Quantification ». Dans ASME 2007 International Mechanical Engineering Congress and Exposition. ASMEDC, 2007. http://dx.doi.org/10.1115/imece2007-43058.

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Real-time quantitative Polymerase Chain Reaction (PCR) is an extremely sensitive and reliable method for quantifying gene expression, allowing subtle shifts in gene expression to be easily monitored. Currently, stationary real-time PCR is readily achieved using fluorescent labels which increase in fluorescence as the DNA is exponentially amplified. Quantitative PCR is used in a myriad of applications. However currently most commercial real-time PCR devices are batch process stationary well based systems, limiting their throughput. Continuous flow microfluidic PCR devices have allowed for advancement in terms of improved PCR throughput and reduced reagent usage. As part of an overall total analysis system a device integrating all the functional steps of continuous flow realtime quantitative PCR has been designed and fabricated. Initially the PCR reaction mixture is segmented into nano-litre PCR reactors which are then thermally cycled on a two temperature fifty cycle flow-through PCR device, which allows laser induced fluorescent imaging of the nanoreactors. Previous studies into continuous flow PCR have demonstrated endpoint fluorescent measurements, however this research allows PCR nanoreactors to be fluorescently monitored after every PCR thermal cycle. Fluorescent optical monitoring is achieved through laser excitation of the nanoreactors while a Charged Coupled Device (CCD) camera is used to record the fluorescent emissions from the nanoreactors. Intensity analysis of the recorded images is then preformed using MATLAB to accurately determine the fluorescence intensity level, thereby allowing real-time quantitative amplification curves to be generated. This has major advantages over existing continuous flow PCR devices which use endpoint fluorescence and capillary electrophoresis, as the amplification curves allow far more information to be gleaned and allow the initial DNA template concentration to be accurately determined.
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« Detection of Gene Expression from Vitis by Real Time Quantitative RT-PCR ». Dans 2018 4th World Conference on Control, Electronics and Computer Engineering. Francis Academic Press, 2018. http://dx.doi.org/10.25236/wccece.2018.15.

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Taha, Aza, Katan Ali et Furat Sabeer. « Quantitation assay of hepatitis C virus RNA using real-time PCR technique ». Dans 4th International Scientific Conference of Cihan University-Erbil on Biological Sciences. Cihan University-Erbil, 2017. http://dx.doi.org/10.24086/bios17.22.

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Huai, Yahong, et Shangzhong Xu. « Real-time Fluorescence Quantitative PCR and Prediction of Bovine Trf 2 Protein Funtion ». Dans International Conference on Biomedical and Biological Engineering. Paris, France : Atlantis Press, 2016. http://dx.doi.org/10.2991/bbe-16.2016.30.

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Botteldoorn, N., H. Werbrouck, N. Rijpens, E. van Coillie, M. Heyndrickx et L. Herman. « Expression study by real-time quantitative RT-PCR of the Salmonella typhimurium mntH gene ». Dans Second International Symposium on Epidemiology and Control of Salmonella in Pork. Iowa State University, Digital Press, 2003. http://dx.doi.org/10.31274/safepork-180809-463.

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Rodríguez-Lázaro, D., M. Pla, M. Scortti, H. J. Monzó et J. A. Vázquez-Boland. « An Optimised Quantitative Real-Time PCR Assay for Listeria Monocytogenes Including and Internal Amplification Control ». Dans 13th World Congress of Food Science & Technology. Les Ulis, France : EDP Sciences, 2006. http://dx.doi.org/10.1051/iufost:20060642.

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Rapports d'organisations sur le sujet "PCR real time quantitativa"

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Harris, D. L. Hank, Isabel Turney Harris, James S. Dickson, Stephen Gaul, Brad T. Bosworth et Lori Feldmann. Quantitative Real-time PCR (qPCR) for the Determination of Salmonella Levels in Lairage. Ames (Iowa) : Iowa State University, janvier 2009. http://dx.doi.org/10.31274/ans_air-180814-1009.

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Zhelev, Doncho V., Christopher Dupuis, Suelynn Ren, Anna Le, Mia Hunt et Henry Gibbons. Single Nucleotide Polymorphisms (SNP)-specific Quantitative Real Time Polymerase Chain Reaction (PCR) Assay for Analyzing Competition and Emergence of the Military Hypersporulating Strains of Bacillus Atrophaeous var. Globigii. Fort Belvoir, VA : Defense Technical Information Center, septembre 2012. http://dx.doi.org/10.21236/ada570597.

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Dilcheva, Valeria, Ivelin Vladov et Svetlozara Petkova. A Comparative Study of Six Trichinella Species by Real-time PCR Assay. "Prof. Marin Drinov" Publishing House of Bulgarian Academy of Sciences, janvier 2018. http://dx.doi.org/10.7546/crabs.2018.01.08.

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Dilcheva, Valeria, Ivelin Vladov et Svetlozara Petkova. A Comparative Study of Six Trichinella Species by Real-time PCR Assay. "Prof. Marin Drinov" Publishing House of Bulgarian Academy of Sciences, janvier 2018. http://dx.doi.org/10.7546/grabs2018.1.08.

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McAvin, James C., et Carl J. Mason. Pre-Clinical Testing of a Real-Time PCR Assay for Diahhreal Disease Agent Cryptosporidium. Fort Belvoir, VA : Defense Technical Information Center, mai 2014. http://dx.doi.org/10.21236/ada600722.

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Bonab, Zahra Hojjati, Parisa Mohammadi, Ezzat Asgarani et Nassim Ghorbanmehr. The Evaluation of Nitrogen Fixation Activity of Soil Cyanobacteria via Reduction Assay and Real-time PCR. "Prof. Marin Drinov" Publishing House of Bulgarian Academy of Sciences, juin 2019. http://dx.doi.org/10.7546/crabs.2019.06.07.

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Faris, Gregory W. Development of Laser-Mediated Nanodroplet Real-Time PCR on Circulating Tumor Cells (CTC) by Microfilter Platform. Fort Belvoir, VA : Defense Technical Information Center, juin 2015. http://dx.doi.org/10.21236/ada621341.

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McAvin, James C., et Carl J. Mason. Norovirus Real Time RT-PCR Detection Technology Transition to the Joint Biological Identification and Diagnosis System (JBAIDS). Fort Belvoir, VA : Defense Technical Information Center, septembre 2012. http://dx.doi.org/10.21236/ada568257.

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McAvin, James C., et Carl J. Mason. Pre-Clinical Testing of Real-Time PCR Assays for Diarrheal Disease Agents of Genera Escherichia and Shigella. Fort Belvoir, VA : Defense Technical Information Center, mai 2014. http://dx.doi.org/10.21236/ada600976.

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Hutchison, Janine R., Gregory F. Piepel, Brett G. Amidan, Michael A. Sydor et Brooke L. Deatherage Kaiser. False Negative Rates of a Macrofoam-Swab Sampling Method with Low Surface Concentrations of Two Bacillus anthracis Surrogates via Real-Time PCR. Office of Scientific and Technical Information (OSTI), mai 2015. http://dx.doi.org/10.2172/1186982.

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