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Periódicos Brasileiros em Medicina Veterinária e Zootecnia

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Development and validation of an efficient real-time quantitative reverse transcription polymerase chain reaction assay of Chinese goose CD4 and CD8

Zhao, QiurongChen, ShunLiu, FeiQi, YulinWang, MingshuJia, RenyongZhu, DekangLiu, MafengCheng, Anchun

Background: CD4+ T cells, which are often referred as T-helper cells, play a central role through secreting various cytokines to enhance immune defense to pathogen. CD8+ T cells, which are called cytotoxic T lymphocytes (CTLs), provide potent defenses against virus infection and intracellular pathogens by killing the targets cells directly. In our previous researches, the conventional and semi-quantitative PCR were used to detect the goose CD4 and CD8. However, the semi-quantitative RT-PCR only detect the relative amount of gene transcription. Quantitative PCR assay was more sensitive than conventional PCR assay, and quantitative PCR assay has a lower limit of sensitivity.Materials, Methods & Results: Contrast to conventional assays, the detection of amplicons by quantitative RT-PCR could be visualized as the amplification progressed. This effect has provided a great deal of insight into the kinetics of the reaction and it is the foundation of kinetic of real-time qPCR. The analysis of gene transcription by qPCR has proven to be an attractive method due to its potential for increasing laboratory throughput, simultaneous processing of several samples as well as more reliable instrumentation. With those in mind, the real-time quantitative reverse transcription PCR (qRTPCR) methods for the detection of goose CD4 and CD8 transcripts were reported here for the first time. With

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