中国兽医科学2017，Vol.47Issue(9)：1140-1144,5.DOI:10.16656/j.issn.1673-4696.2017.09.012
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
摘要
Abstract
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 bpfragment of CanineCV,and the other for amplification of a 668 bpfragment 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/μ L for CanineCV DNA and 312 copies/μL 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PCRKey words
canine circovirus/canine parvovirus/duplex PCR分类
农业科技引用本文复制引用
曹亮,张金勇,钱爱东,金宁一,孙文超,鲁会军,田明尧,刘云霞,谢长占,赵冠宇,韩继成,肖鹏鹏..犬圆环病毒及犬细小病毒双重PCR检测方法的建立[J].中国兽医科学,2017,47(9):1140-1144,5.基金项目
国家自然科学基金资助项目(31272573) （31272573）
