套式RT-PCR检测猪繁殖和呼吸综合征病毒的研究  

RESEARCH ON THE TESTING OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS BY NESTRT-PCR

作　　者：赵耘[1] 张广州[1] 秦玉明[1] 宁宜宝[1] 王琴[1] 

机构地区：[1]中国兽医药品监察所,北京100081

出　　处：《微生物学通报》2003年第6期77-80,共4页Microbiology China

基　　金：国家973计划资助项目(No.G1999011905)

摘　　要：参照ATCC VR-2332株及LV株保守区段设计了3条引物,以此建立了检测猪繁殖和呼吸综合征病毒的套式RT-PCR方法。利用其分别对ATCC VR-2332株、LV株及B13株进行套式RT-PCR,结果从3个不同地区分离的毒株中均能特异性的扩增出相应的片段,大小分别约为430bp(预期片段为430bp)、410bp(预期片段为413bp)及410bp(预期片段为413bP),而3个非PRRSV的病毒(猪瘟病毒、细小病毒及伪狂犬病毒)均未扩增出相应的片段。其敏感性可达到10^(-2)TCID_(50),比一步法RT-PCR方法敏感性提高了10000倍。本方法的建立使猪繁殖和呼吸综合征病毒的检测更为敏感、快捷及准确。Three primer were designed based on the consevered area of the genetic of the ATCC VR-2332 strain and LV strain. And the nest RT-PCR of testing porcine reproductive and respiratory syndrome virus were developed. The nest RT-PCR against ATCC VR-2332 strain, LV strain and B13 strain were done by this method.The DNA fragment were obtained specially from the three strains isolated from different region. The size were 430bp (430bp) , 410bp (413bp) and 410 bp (413bp) separately. But the DNA fragment were not obtained from HCV, PPV and PRV. Its sensitivity was 10-2 TCID50. It's sensitivity increased 10000 times than one step RT-PCR. It should make the method of testing porcine reproductive and respiratory syndrome virus more sensitive, fast and accurate.

关 键 词：套式RT-PCR 特异性 敏感性 一步法RT-PCR 

分 类 号：S852.65[农业科学—基础兽医学]