鸡传染性支气管炎病毒793/B RT-PCR检测方法的建立及应用

Establishment and application of RT-PCR technigue for detection of the 793/B serotype of infectious bronchitis virus

根据GenBank已发表的鸡传染性支气管炎病毒(Infectious bronchitis virus,IBV)793/BS1基因序列,设计1对引物,扩增891bp特异性核酸片段,建立了检测IBV793/B的RT-PCR方法。特异性试验结果表明,IBV793/B能扩增出891bp的核酸片段,而IBVM41、H120、H52毒株以及新城疫病毒(NDV)、禽流感病毒(AIV)、传染性法氏囊病病毒(IBDV)均无特异性条带出现。敏感性试验结果表明,该方法的最低检出量为10pg的模板。上述结果表明,本试验所建立的RT-PCR方法敏感性高、特异性强。利用建立的RT-PCR方法对从山东省分离的8株疑似鸡IBV793/B进行检测,结果7株为阳性。该方法的建立为IBV793/B的诊断及流行病学调查提供了可靠的方法。

According to the sequences of the 793/B serotype of IBV S1 gene published in GenBank,a pair of primers was designed and synthesized to specifically amplify the 891 bp nucleotide fragment,the RT-PCR technique for detection of the 793/B serotype of IBV was established. The 891 bp special strip was found only in the PCR amplification of the 793/B serotype of IBV in the specificity assay,there were no the same strip appeared in the other viruses and strains of IBV. The sensitivity result indicated that as little as 10 pg in sample were detected in PCR. Seven of the eight uncertain isolates detected by the established RT-PCR technique were positive. The above resuits showed that the technique provided a more sensitive,specific and rapid method for diagnosis and epidemiological survey of the 793/B serotype of IBV.