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Báo cáo y học: " The development of a rapid SYBR one step real-time RT-PCR for detection of porcine reproductive and respiratory syndrome virus"

The development of a rapid SYBR one step real-time RT-PCR for detection of porcine reproductive and respiratory syndrome virus | Tian et al. Virology Journal 2010 7 90 http www.virologyj.eom content 7 1 90 VIROLOGY JOURNAL SHORT REPORT Open Access The development of a rapid SYBR one step real-time RT-PCR for detection of porcine reproductive and respiratory syndrome virus Hong Tian JingYan Wu YouJun Shang Yan Cheng and XiangTao Liu Abstract Background Prompt detection of PRRSV in the field samples is important for effective PRRS control thereby reducing the potentially serious economic damage which can result from an outbreak. In this study a rapid SYBR-based one step real-time RT-PCR quantitative reverse transcription PCR qRT-PCR has been developed for the detection of porcine reproductive and respiratory syndrome virus PRRSV . Primers were designed based on the sequence of highly conservative region of PRRSV N gene. Results The sensitivity of the real-time qRT-PCR assay was achieved through PRRSV ch-1a RNA for the generation of a standard curve. The detection limit of the assay was found to be 9.6 RNA copies per reaction mixture. This assay had excellent intra- and inter-assay reproducibility as in total 65 field samples were screened for the presence of PRRSV by conventional RT-PCR in parallel with qRT-PCR and the detection rate increased from 60.0 to 76.9 . Moreover the specificity result indicated that this assay could reliably differentiate PRRSV from the other swine viral diseases such as classical swine fever virus CSFV swine vesicular disease virus SVDV and vesicular exanthema of swine virus VESV . Conclusion The real-time qRT-PCR assay described in this report allows the rapid specific and sensitive laboratory detection of PRRSV in field samples. Background Porcine reproductive and respiratory syndrome virus PRRSV is a member of the family Arteriviridae 1 and it was characterized by respiratory disease in young pigs and severe reproductive failure in sows including abortion stillbirths and weak piglets 2 . PRRS has caused immense economic losses in the pig industry and is .