A型猪流感病毒实时荧光定量PCR检测方法的建立及应用  

Establishment and Application of a Real-time Quantitative PCR Method for Rapid Detection of Swine Influenza Virus A-type

作　　者：欧阳振宇[1] 唐连飞[1] 朱中武[1] 肖家勇[1] 孟芳[1] 黄萍[1] 吴愈柏[2] 

机构地区：[1]湖南出入境检验检疫局检验检疫技术中心,长沙410004 [2]湖南出入境检验检疫局动物检疫处,长沙410004

出　　处：《中国畜牧兽医》2010年第9期75-77,共3页China Animal Husbandry & Veterinary Medicine

基　　金：国家质量监督检验检疫总局资助项目(2007IK011)

摘　　要：根据WHO推荐的检测A型猪流感病毒的引物和探针序列,建立了A型猪流感病毒实时荧光定量PCR检测方法。使用含有选定检测序列的重组质粒标准品绘制标准曲线,检测结果表明,该方法的敏感性可达100拷贝/25μL反应体系,显示出良好的敏感性和重复性,临床上已用于实验室鼻拭子样品的检测,并成为一种快速定量检测A型猪流感病毒的方法。The specific primers and probe were designed according to the region of genes of swine influenza virus A-type that recommend by WHO.A real-time fluorescent quantitative PCR method was established for detection of swine influenza virus A-type.Series of dilutions of recombinant plasmids were prepared and used to generate standard curves.The results showed that the real-time fluorescent quantitatine PCR was capable of detecting 100 copies of swine influenza virus A-type genes per 25 μL reaction system.The method was very sensitive and reprducibilitive,had been used for clinical laboratory testing of nasal swab samples,and could be used for rapid quantitative detection of swine influenza virus A-type.

关 键 词：A型猪流感病毒 实时荧光定量PCR H1N1亚型 

分 类 号：S852.659.5[农业科学—基础兽医学]