应用RT-nested PCR检测猪传染性胃肠炎病毒

Study on detecting transmissible gastroenteritis virus by reverse transcription-nested polymerase chain reaction

根据GenBank中已发表的猪传染性胃肠炎病毒(TGEV)基因序列,利用Primer5.0程序软件,设计并合成了M基因的2对引物,以mRNA为模板,通过RT-PCR,RT-nestedPCR方法,成功扩增出长度为1328bp和1037bp的TGEV目的片段,但未扩增出猪传染性腹泻病毒(PEDV)片段。在优化RT-PCR反应条件的基础上,建立了快速检测TGEV的诊断方法。结果表明,RT-nestedPCR方法可用于检测猪传染性胃肠炎病毒,而且此方法简单省时、灵敏性高,可以作为检测RNA病毒的一种分子生物学方法。

Two pairs of oligonucleotide primers appropriate for PCR amplication were selected based on the gene encoding of the M protein of TGEV.The amplified products of TGEV were 1 328 bp,1 037 bp respectively,but as to PEDV nothing was amplified.By a reverse transcription-nested polymerase chain reaction (RT-nested PCR) assay,a quick detection of TGEV after optimization of the reaction conditions was developed.The results showed that RT-PCR assay can be used to detect the virus of RNA.