猪流感病毒RT-PCR检测方法的建立及初步应用

The Establishment and Preliminary Application of a RT-PCR for Detection of Swine Lnfluenza Virus

摘要: 根据GenBank发表H1N1、H1N2和H3N2亚型猪流感病毒M基因片段的引物序列, 设计合成1对引物, 建立猪流感病毒RT-PCR检测方法。结果表明:建立的方法能扩增出675bp的基因片段, 同条件扩增伪狂犬病毒、猪瘟病毒、圆环病毒Ⅱ型及猪繁殖与呼吸综合征病毒均为阴性;该方法可检测到3.5pg猪流感的核酸, 能从被流感病毒感染的小白鼠和疑似猪流感的病料中检测到猪流感病毒。结果表明, 建立的猪流感病毒RT-PCR方法具有特异性强、敏感性高和重复性好等特点, 适用于对H1N1、H1N2和H3N2亚型猪流感病毒的快速诊断。

Abstract: A pair of primers was designed based on the M gene sequence of the Swine influenza virus (SIV) published in Genbank, and the matched RT-PCR assay was developed.The 675 bp specific fragment was amplified from H1N1, H1N2 and H3N2subtype Swine influenza viruses.Negative results were found on PRV, CSFV, PCV2 and PRRSV.The developed method was sensitive to detect RNA as little as 3.5pg, and Siv could be detected directly from lungs of infected mice and pathogenic swine.This RT-PCR assay was proved to be specific, sensitive, reproducible and suitable for rapid diagnosis of H1N1, H1N2 and H3N2.