检测羊肠道病毒RT-PCR方法的建立及初步应用  

Establishment and preliminary application of RT-PCR method for the detection of caprine/ovine enterovirus

作　　者：董坤 胡俊英 李卓宸 蔡梦露 章凡 王玮玉 王浴光 王新平 DONG Kun;HU Junying;LI Zhuochen;CAI Menglu;ZHANG Fan;WANG Weiyu;WANG Yu-guang;WANG Xinping(Key Laboratory for Zoonosis,the Ministry of Education,College of Veterinary Medicine,Jilin University,Changchun 130062,China)

机构地区：[1]吉林大学动物医学学院教育部人兽共患病研究重点实验室,吉林长春130062

出　　处：《中国兽医学报》2021年第10期1949-1952,共4页Chinese Journal of Veterinary Science

基　　金：“十三五”国家重点研发计划资助项目(2016YFD0500904,2017YFD0500104)。

摘　　要：羊肠道病毒(caprine enterovirus,CEV)感染为国内外新发疫病,其诊断方法尚缺乏研究。本研究依据CEV-JL14毒株的基因组序列,设计合成了1对特异性引物,建立了检测CEV的RT-PCR方法,并对其特异性、敏感性和重复性进行测定,同时应用该方法对疑似CEV感染的临床样品进行检测。结果显示,建立的检测CEV感染的RT-PCR方法具有特异性好、敏感性高、快速和简便等特点,最低检出限为1.13×10^(3)copies/μL。以建立的RT-PCR方法对通过ELISA方法确定的6份阳性样品和22份阴性样品进行检测,结果显示两者的符合率为100%。本研究建立的RT-PCR方法可用于CEV感染的诊断和流行病学调查。Caprine enterovirus(CEV)infection is an emerging disease at home and abroad that causes a significant economic loss to goat/sheep industry.Currently,few methods are available for diagnosis of this infection.In this study,a pair of goat enterovirus-specific primers was designed and synthesized based on the genome sequence of goat enterovirus CEV-JL14 strain.RT-PCR method for the detection of goat enterovirus was established,and the specificity,sensitivity and repeatability were tested.In addition,the established RT-PCR methods was compared with a sandwich ELISA for the detection of clinical samples suspected of CEV infection.The results showed that the established RT-PCR method was specific for detecting CEV with a detection sensitivity of 1.13×10^(3)copies/μL.The established RT-PCR method was used to detect six positive samples and 22 negative samples determined by ELISA method,and the coincidence rate of the two methods was 100%.The above results indicate that the established RT-PCR method can be used for the diagnosis and epidemiological investigation of CEV infection.

关 键 词：羊肠道病毒 EV-G20 反转录-聚合酶链式反应 

分 类 号：S852.65[农业科学—基础兽医学]