鸡细小病毒半巢式PCR检测方法的建立及初步应用

发布时间：2018-06-05 18:16

本文选题：鸡细小病毒 + 半巢式PCR　；参考：《中国畜牧兽医》2017年05期

【摘要】：为建立一种快速、特异、灵敏的检测鸡细小病毒(chicken parvovirus,ChPV)的方法,根据ChPV的保守基因NS1设计了3条特异性引物,建立并优化了能快速检测ChPV的半巢式PCR方法,对其进行特异性和敏感性试验,并用所建立的方法对48份临床样品进行了检测。特异性和敏感性试验结果显示,建立的半巢式PCR只对ChPV敏感,扩增产物为186bp的特异性条带;其最低能检测到5.62fg/μL的ChPV DNA;而对鸡新城疫病毒、H9亚型禽流感病毒、马立克氏病病毒、鸡传染性喉气管炎病毒、鸡传染性支气管炎病毒不敏感。临床检测结果显示,同时对48份临床样品进行检测,检出率为16.67%(8/48),提示广西区内鸡群存在ChPV感染。本研究建立的ChPV半巢式PCR方法适用于ChPV的临床检测。
[Abstract]:In order to establish a rapid, specific and sensitive method for the detection of chicken parvovirus (ChPV), three specific primers were designed according to the conserved gene NS1 of ChPV, and a semi-nested PCR method was established and optimized for rapid detection of ChPV. The specificity and sensitivity were tested and 48 clinical samples were detected by the established method. The results of specificity and sensitivity tests showed that the semi-nested PCR was only sensitive to ChPVs, and the amplified product was a specific band of 186bp. The lowest 5.62fg/ 渭 L 5.62fg/ DNA could be detected, while the H9 subtype avian influenza virus of Newcastle disease virus, Marek's disease virus, was detected. Chicken infectious laryngotracheitis virus, chicken infectious bronchitis virus is insensitive. The results of clinical examination showed that 48 clinical samples were detected and the detection rate was 16.67% / 48%, indicating that ChPV infection existed in chickens in Guangxi. The semi-nested PCR method developed in this study is suitable for clinical detection of ChPV.
【作者单位】：广西壮族自治区兽医研究所广西兽医生物技术重点实验室;南宁市良凤农牧有限责任公司;
【基金】：广西科技项目(15-140-31-A-1、15-140-31-B-1、桂科AB16380003) 南宁市科技攻关项目(20152308) 国家“万人计划”领军人才专项(2016-37-88)
【分类号】：S858.31

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