实验动物与比较医学2017，Vol.37Issue(1)：32-35,4.DOI:10.3969/j.issn.1674-5817.2017.01.007
大鼠细小病毒双重PCR检测方法的建立
Development of Dual PCR for Detection of Rat Parvovirus
摘要
Abstract
Objective To establish a PCR method for rapidly detecting rat parvovirus.Methods According to the characters of nucleotide sequence of rat parvovirus,a dual PCR was developed to differentially detect RPV and other three rat parvovirus (H-1/KRV/RMV).A pair of primers amplifying VP2 gene were applied to exclusively amplify RPV while another pair of primers targeting the NS 1 gene was designed for specific amplification of H-1,KRV and RMV primer.Result Rat parvovirus were screened without detecting minute virus of mice which had high nucleotide homology with rat parvovirus and also negative for three other microorganism pathogen.Sensitivity tests showed that the minimum detectable concentration was as low as 1 000 copies μL.When dual PCR combined with sequencing were applied to detect 23 clinical samples,seven samples detected were RMV positive including one coinfected with RPV.Conclusion The dual PCR was verified to be specific and sensitive which could be used as a reliable method to screen rat parvovirus.关键词
大鼠细小病毒(RPV)/H-1/KRV/RMV/双重PCRKey words
Rat Parvovirus (RPV)/H-1/KRV/RMV/Dual PCR分类
农业科技引用本文复制引用
饶丹,徐凤娇,刘向楠,刘助红,黄韧,张钰,郭鹏举,朱余军,伍妙梨,袁文,王静,尹雪琴,丛锋,练月晓,黄碧洪..大鼠细小病毒双重PCR检测方法的建立[J].实验动物与比较医学,2017,37(1):32-35,4.基金项目
广东省科技计划项目(2014B070706006) （2014B070706006）
国家科技支撑计划(2015BAI07B00)广东省科技计划项目(2013B060400028) （2015BAI07B00）
