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猪细小病毒和猪源脑心肌炎病毒的二重PCR检测方法的建立
引用本文:朱向博,刘业兵,王晶钰,张晓杰,梁耀峰,王利勤,董睿,陈婷,李成山.猪细小病毒和猪源脑心肌炎病毒的二重PCR检测方法的建立[J].中国畜牧兽医,2012,39(8):31-35.
作者姓名:朱向博  刘业兵  王晶钰  张晓杰  梁耀峰  王利勤  董睿  陈婷  李成山
作者单位:1. 西北农林科技大学动物医学院,陕西杨凌712100;中国兽医药品监察所,北京100081
2. 中国兽医药品监察所,北京,100081
3. 西北农林科技大学动物医学院,陕西杨凌,712100
基金项目:国家自然科学基金项目,十一五国家科技支撑计划项目
摘    要:试验旨在建立能同时检测猪细小病毒(porcine parvovirus,PPV)和猪源脑心肌炎病毒(encephalomyocarditis virus,EMCV)的二重PCR检测方法。根据GenBank中PPV VP2基因和EMCV 3D基因序列,利用Premier 5.0软件设计出PPV和EMCV的特异性引物。通过优化反应条件,建立了能同时特异性检测PPV和EMCV的二重PCR方法,PPV和EMCV的敏感性检测极限分别达7.62和1.22×10-1 pg/μL。通过对临床36份样品的检测结果表明,该二重PCR检测方法敏感、特异,适合于临床检测应用。

关 键 词:PPV  EMCV  猪源脑心肌炎病毒  二重PCR  
收稿时间:2012-01-09

Establishment of a Diplex PCR Assay for Detection of PPV and Porcine EMCV
ZHU Xiang-bo , LIU Ye-bing , WANG Jing-yu , ZHANG Xiao-jie , LIANG Yao-feng , WANG Li-qin , DONG Rui , CHEN Ting , LI Cheng-shan.Establishment of a Diplex PCR Assay for Detection of PPV and Porcine EMCV[J].China Animal Husbandry & Veterinary Medicine,2012,39(8):31-35.
Authors:ZHU Xiang-bo  LIU Ye-bing  WANG Jing-yu  ZHANG Xiao-jie  LIANG Yao-feng  WANG Li-qin  DONG Rui  CHEN Ting  LI Cheng-shan
Institution:1. College of Veterinary Medicine,Northwest A & F University,Yangling 712100,China; 2. China Institute of Veterinary Drug Control,Beijing 100081,China
Abstract:Establish a diplex PCR assay was to detect porcine parvovirus (PPV) and porcine encephalo myocarditis virus (EMCV) simultaneously.According to PPV VP2 gene sequences and EMCV 3D gene sequences in GenBank, special primers of PPV and EMCV were designed by Premier 5.0 software. Trough optimizing the reaction conditions, a diplex PCR assay,which could detect PPV and EMCV simultaneously,was established. The sensitivity of PPV and EMCV were 7.62 and 1.22×10-1 pg/μL respectively. Clinical tests of 36 samples showed that the diplex PCR detection assay was sensitive, specific, and suitable for clinical application.
Keywords:PPV  EMCV  porcine encephalomyocarditis virus  diplex PCR assay
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