摘要
为建立一种检测内蒙古地区绵羊梅迪-维斯纳病毒(MVV)的TaqMan荧光定量PCR方法,根据高度保守的gag基因序列设计特异性引物和探针,通过对反应条件和反应体系的优化,建立了快速检测MVV的TaqMan实时荧光定量PCR方法。结果显示,扩增相关系数为0.999,对重组标准质粒的最低检测量为1.0×100 copies/μL,对羊的其他肺炎表现常见病原核酸无扩增反应;利用该方法对内蒙古地区12份疑似病例的肺组织进行检测,其中3份为MVV阳性。结果表明,本研究建立的实时荧光定量PCR方法在MVV的快速、准确诊断中具有良好的应用前景,也将为内蒙古地区防控梅迪-维斯纳病提供可靠的检测手段。
In order to detect Maedi-Visna virus(MVV) in Inner Mongolia,we designed a pair of primers and a probe based on the highly conserved gag gene of MVV. By optimizing the reaction conditions and reaction system,a TaqMan real-time PCR method for rapid detection of MVV was established. The results indicated that it had a good linear relationship(R2=0.999),and there was no cross-reaction with other nucleic acids of common pathogens of ovine pneumonia.The lowest detection limit was 1.0×100 copies/μL.Finally,this method was applied to examine the lung tissues of 12 suspected cases from Inner Mongolia,and 3 out of 12 samples were MVV positive. Therefore,the established method exhibits a good application prospect for the rapid and accurate diagnosis of MVV,and it will be a reliable detection method for the prevention and control of Maedi-Visna disease in Inner Mongolia.
作者
李慧萍
张良
徐斯日古楞
王多智
刘淑英
LI Hui-ping;ZHANG Liang;Xusiriguleng;WANG Duo-zhi;LIU Shu-ying(College of Veterinary Medicine,Inner Mongolia A gricultural University,Hohhot 010018,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2021年第7期828-833,共6页
Chinese Veterinary Science
基金
国家自然科学基金项目(31760721,32072819)
内蒙古草原英才创新团队项目(20151031)
内蒙古应用研究项目(2019GG240)。
