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A multiplex RT-PCR assay for rapid and simultaneous detection of four RNA viruses in swine

文献类型： 外文期刊

作者： Zhao, Yan 1 ; Liu, Feifei 1 ; Li, Qingmei 2 ; Wu, Mengfan 1 ; Lei, Lei 1 ; Pan, Zishu 1 ;

作者机构： 1.Wuhan Univ, Coll Life Sci, Modern Virol Res Ctr, State Key Lab Virol, Wuhan 430072, Peoples R China

2.Henan Acad Agr Sci, Key Lab Anim Immunol, Zhengzhou 450002, Henan, Peoples R China

关键词： Multiplex RT-PCR; Simultaneous detection; Virus infection; Swine

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）

ISSN： 0166-0934

年卷期： 2019 年 269 卷

页码：

收录情况： SCI

摘要： A multiplex reverse transcription polymerase chain rection (mRT-PCR) was developed for simultaneous detection of four RNA viruses in swine. The conserved target sequences directed to classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis coronavirus (TGEV) were selected based on alignments of genomic sequences and then specific primers were designed. The mRT-PCR assay was developed and evaluated for its specificity and sensitivity. The expected product from the single viral template was amplified by mRT-PCR and no spurious PCR amplification occurred from the genomic RNA or DNA of other pathogens. For single virus or different combinations of two viruses the detection limit of mRT-PCR was consistent with a single RT-PCR wtith 1 x 10(3) copies. For different combinations of the three viruses or four viruses, sensitivity of PEDV detection partially decreased. All of positive clinical specimens by the mRT-PCR were identically confirmed using Taqman RT-qPCR. Therefore, the mRT-PCR is a useful tool for epidemiological studies and laboratory diagnosis of single virus and/or mixed infections in swine.

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