犬圆环病毒及犬细小病毒双重PCR检测方法的建立  

Establishment of duplex PCR for the detection of canine circovirus and canine parvovirus

作　　者：曹亮[1,2] 孙文超[2] 鲁会军[2] 田明尧[2] 刘云霞[1,2] 谢长占 赵冠宇[2] 韩继成[2] 肖鹏鹏 张金勇[1,2] 钱爱东[1] 金宁一[1,2] 

机构地区：[1]吉林农业大学动物科学技术学院,吉林长春130118 [2]军事医学科学院军事兽医研究所,吉林长春130122

出　　处：《中国兽医科学》2017年第9期1140-1144,共5页Chinese Veterinary Science

基　　金：国家自然科学基金资助项目(31272573)

摘　　要：为建立能同时检测犬圆环病毒(CanineCV)和犬细小病毒(CPV)的方法,根据CanineCV和CPV基因组的保守区域设计2对特异性引物,预期特异性扩增CanineCV和CPV的片段大小分别为984和668 bp。并将反应条件进行优化,建立了CanineCV和CPV的双重PCR检测方法。特异性检测结果显示,该方法能特异性扩增CanineCV和CPV的目的条带,且未扩增出其他犬胃肠道病原;CanineCV和CPV的最低检出限分别为85.7和312 copies/L。对采集自广西地区的223份临床样品的检测结果显示,CanineCV的阳性率为2.24%(5/223)、CPV的阳性率为5.38%(12/223),后者与常规PCR检测方法相一致。结果表明,建立的双重PCR方法可以用于CanineCV和CPV的检测和流行病学调查。To develop a duplex PCR assay for the detection and differentiation of canine circovirus （CanineCV） and canine parvovirus （CPV）,two pairs of specific primers were designed according to the genome conservative sequence of CanineCV and CPV,of which one was used for specifically amplification of a 984 bp fragment of CanineCV,and the other for amplification of a 668 bp fragment of CPV.The reaction conditions of the annealing temperature and the primer concentration were optimized.Specificity test showed no amplification was observed when other common pathogens for canine gastrointestinal diseases were used as template.The detection limits of assay were 85.7 copies/L for CanineCV DNA and 312 copies/L for CPV DNA.Furthermore,the established assay was used to detect clinical samples collected from Guangxi Zhuang Autonomous Region.The results showed that the positive rates of CanineCV and CPV were 2.24%（5/223） and 5.38%（12/223）,respectively,and the latter was fully corresponding with that of the conventional PCR assay.These results suggested that the developed duplex PCR assay was suitable for clinical detection and epidemiology surveillance of CanineCV and CPV.

关 键 词：犬圆环病毒 犬细小病毒 双重PCR 

分 类 号：S852.659.2[农业科学—基础兽医学]