摘要
旨在建立鸡细小病毒(ChPV)与鸡新城疫病毒(NDV)双重PCR检测方法,参考ChPV NS1基因和NDV F基因的保守区域,设计筛选、合成验证两对ChPV和NDV特异性引物,通过优化其反应体系,以及双重PCR的特异性、敏感性试验来评估建立的ChPV与NDV双重PCR检测方法。结果表明,建立的双重PCR同时扩增出687 bp ChPV和453 bp NDV片段,对ChPV与NDV的敏感性分别达到64 fg和25 fg,但对其他对照组病原体均无扩增,对ChPV与NDV混合感染的临床病料的检测结果与各病毒单项PCR检测结果相符。说明建立的ChPV与NDV双重PCR检测方法可用于临床上两种病毒混合感染的快速诊断。
A duplex PCR assay for detecting Chicken parvovirus(ChPV)and Newcastle disease virus(NDV)was developed to provide technical support for effective control of ChPV and NDV.According to the conserved gene sequences of ChPV NS1 and NDV F in GenBank,two sets of specific primers were designed.The duplex PCR for simultaneous detection of ChPV and NDV was established by optimizing the reaction conditions.The result indicated that the duplex PCR assay successfully amplified 687 bp for ChPV,453 bp for NDV.The sensitivity of the assay was 64 fg for ChPV,25 fg for NDV,but it was not sensitive to other chicken pathogens,such as MDV,ARV,APV,IBV,AILTV and E.coli.The results detected by the duplex PCR had a 100%coincidence with the results detected by single PCR for the ChPV and NDV respectively.The duplex PCR assay is a quick,sensitive and specific test for detection of ChPV and NDV.
作者
奉彬
谢芝勋
邓显文
张艳芳
谢丽基
谢志勤
范晴
曾婷婷
罗思思
黄娇玲
王盛
刘加波
FENG Bin;XIE Zhi-xun;DENG Xian-wen;ZHANG Yan-fang;XIE Li-ji;XIE Zhi-qin;FAN Qing;ZENG Ting-ting;LUO Si-si;HUANG Jiao-ling;WANG Sheng;LIU Jia-bo(Lingui Center for Animal Disease Control and Prevention,Lingui,Guangxi,541199,China;Guangxi Veterinary Research Institute,Guangxi Key Laboratory of Veterinary Biotechnology,Nanning,Guangxi,530001,China)
出处
《动物医学进展》
北大核心
2019年第10期1-5,共5页
Progress In Veterinary Medicine
基金
广西科技项目(AD17195083,AA17204057)
“广西八桂学者”专项(2019-79)
国家“万人计划”领军人才专项(W02060083)
