| 大鼠细小病毒双重PCR检测方法的建立 |
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| 引用本文： | 饶丹,朱余军,伍妙梨,袁文,王静,尹雪琴,丛锋,练月晓,黄碧洪,徐凤娇,刘向楠,刘助红,黄韧,张钰,郭鹏举. 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, 0(1): 32-35. DOI: 10.3969/j.issn.1674-5817.2017.01.007 |
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| 作者姓名： | 饶丹 朱余军 伍妙梨 袁文 王静 尹雪琴 丛锋 练月晓 黄碧洪 徐凤娇 刘向楠 刘助红 黄韧 张钰 郭鹏举 |
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| 作者单位： | 广东省实验动物监测所,广州,510663 |
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| 基金项目： | 广东省科技计划项目(2014B070706006)，国家科技支撑计划(2015BAI07B00)广东省科技计划项目(2013B060400028) |
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| 摘 要： | 目的 建立快速检测大鼠细小病毒的PCR方法.方法 根据大鼠细小病毒基因序列的特点,建立鉴别检测大鼠细小病-(RPV)与其他三种大鼠细小病毒(H-1、KRV、RMV)的双重PCR方法.根据RPV核酸序列的性质,在VP2基因筛选了一对用于特异扩增RPV株的引物,在NS1基因设计了一对用于特异性扩增H-1,KRV和RMV的引物.结果 两对引物组成的二重PCR方法可以特异性的扩增RPV而不扩增核酸同源性高的小鼠细小病毒和多种其他病原微生物;敏感性实验表明,二重PCR的最低检测限可达1 000拷贝/μL.双重PCR结合测序在检测23份临床样本中,检测出7份RMV阳性,包括1份与RPV共感染阳性样本.结论 本研究建立的检测RPV的双重PCR方法具有特异性好、灵敏度高及操作简单等优点,是一种简便、可靠的检测方法.
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| 关 键 词： | 大鼠细小病毒(RPV) H-1 KRV RMV 双重PCR |
Development of Dual PCR for Detection of Rat Parvovirus |
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| RAO Dan,ZHU Yu-jun,WU Miao-li,YUAN Wen,WANG Jing,YIN Xue-qin,CONG Feng,LIAN Yue-xiao,HUANG Bi-hong,XU Feng-jiao,LIU Xiang-nan,LIU Zhu-hong,HUANG Ren,ZHANG Yu,GUO Peng-ju. Development of Dual PCR for Detection of Rat Parvovirus[J]. Laboratory Animal and Comparative Medicine, 2017, 0(1): 32-35. DOI: 10.3969/j.issn.1674-5817.2017.01.007 |
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| Authors: | RAO Dan ZHU Yu-jun WU Miao-li YUAN Wen WANG Jing YIN Xue-qin CONG Feng LIAN Yue-xiao HUANG Bi-hong XU Feng-jiao LIU Xiang-nan LIU Zhu-hong HUANG Ren ZHANG Yu GUO Peng-ju |
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| 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. |
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| Keywords: | Rat Parvovirus (RPV) H-1 KRV RMV Dual PCR |
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