禽流感病毒与鸡细小病毒二重PCR检测方法的建立

摘要

为建立一种能同时鉴别诊断禽流感病毒(avian influenza virus,AIV)和鸡细小病毒(chicken parvovirus,ChPV)的检测方法,本研究根据GenBank中AIV 的M基因和ChPV 的NS基因保守序列,分别设计并筛选出两对特异性引物,用于AIV和ChPV的检测.通过优化反应条件,建立了AIV和ChPV二重PCR检测方法.试验结果表明,该方法特异好,能同时检测AIV和ChPV,对其他常见的禽病病原体均未反应;该法对AIV和ChPV的检测下限均为100 fg;对159份临床样品检测结果与PCR阳性产物测序结果一致.本研究建立的AIV和ChPV二重PCR检测方法具有特异性好、灵敏度高的特点,对 AIV 和ChPV的防制具有重要意义.%In order to establish a method to simultaneously detect avian influenza virus (AIV) and chicken parvovirus (ChPV),two pairs of specific primers were designed according to the sequences of AIV M gene and ChPV NS gene in GenBank.The duplex PCR assay was established by optimizing the reaction conditions.The tests showed that this method had high specificity, could simultaneously detect AIV and ChPV and no specific band was amplified for other subtypes avian pathogenic virus.The sensitivity result showed that the lower detection limit of this method was 100 fg.The results of 159 clinical samples were consistent with the sequencing results of PCR positive product.The double PCR methods for detection of AIV and ChPV established in this study had the characteristics of good specificity and high sensitivity, which was of great significance to the prevention control of AIV and ChPV.