5种猪病毒性传染病病原的多重PCR检测方法的建立

发布时间：2018-06-26 13:46

本文选题：多重PCR + 非洲猪瘟病毒　；参考：《中国预防兽医学报》2017年02期

【摘要】：为建立同时检测非洲猪瘟病毒(ASFV)、水疱性口炎病毒(VSV)、猪口蹄疫病毒(FMDV),猪瘟病毒(CSFV)以及猪伪狂犬病病毒(PRV)的多重RT-PCR检测方法,本研究根据Gen Bank中登录的参考病毒株序列,选择各病毒的保守序列设计5对特异性引物,通过优化反应条件,建立了一种可以同时快速检测以上5种病毒的多重RT-PCR检测方法。结果显示,该方法对不同的细菌或病毒模板扩增结果均为阴性,特异性强;敏感性试验表明该方法对PRV、CSFV、ASFV、VSV和FMDV的核酸最少检出量分别为8.82×10~3拷贝/μL、6.87×10~4拷贝/μL、5.71拷贝/μL、4.93×10~4拷贝/μL和4.32×10~2拷贝/μL。以上结果表明该方法快速、灵敏、特异性强,对以上5种猪病病毒能够进行快速鉴别检测,为其临床诊断与流行病学调查提供了有效的检测方法。
[Abstract]:In order to establish a multiplex RT-PCR method for simultaneous detection of African classical swine fever virus (ASFV), vesicular stomatitis virus (VSV), foot-and-mouth disease virus (FMDV), classical swine fever virus (CSFV) and porcine pseudorabies virus (PRV), a multiplex RT-PCR method was developed for the detection of African swine fever virus (ASFV), vesicular stomatitis virus (VSV), swine foot and mouth disease virus (FMDV), swine fever virus (CSFV) and porcine pseudorabies virus (PRV). Five pairs of specific primers were designed according to the conserved sequence of each virus. By optimizing the reaction conditions, a multiplex RT-PCR method was developed for the simultaneous and rapid detection of the above five viruses. The results showed that the PCR assay was negative and specific for different bacterial or viral templates, and the sensitivity test showed that the minimum detectable amounts of nucleic acid for PRVG CSFV VSV and FMDV were 8.82 脳 10 ~ (-3) copies / 渭 L ~ 6.87 脳 10 ~ (4) copies / 渭 L 5.71 copies / 渭 L 4.93 脳 10 ~ (4) copies / 渭 L and 4.32 脳 10 ~ (2) copies / 渭 L, respectively. The results showed that the method was rapid, sensitive and specific, and could be used for rapid identification and detection of the above five kinds of swine disease viruses, which provided an effective method for clinical diagnosis and epidemiological investigation.
【作者单位】：四川农业大学动物医学院;动物疫病与人类健康四川省重点实验室;
【基金】：国家“十二五”国家科技支撑计划(2013BAD12B04)
【分类号】：S852.651

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