[1]魏文康,吕殿红,温肖会,等.食源性动物组织中CSFV和PRRSV双重双色荧光定量RT-PCR检测方法的建立及应用[J].中国预防兽医学报,2011,(08):620-624.
 WEI Wen-kang,LV Dian-hong,WEN Xiao-hui,et al.Development of the duplex real-time RT-PCR based on dual labeled fluorescent probes for detection of classical swine fever virus and porcine reproductive and respiratory syndrome virus from food-oriented pig tissues[J].Chinese journal of preventive veterinary medicine,2011,(08):620-624.
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食源性动物组织中CSFV和PRRSV双重双色荧光定量RT-PCR检测方法的建立及应用
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2011年08期
页码:
620-624
栏目:
诊断和检测技术
出版日期:
2011-08-15

文章信息/Info

Title:
Development of the duplex real-time RT-PCR based on dual labeled fluorescent probes for detection of classical swine fever virus and porcine reproductive and respiratory syndrome virus from food-oriented pig tissues
作者:
魏文康;吕殿红;温肖会;罗胜军;黄忠;周秀蓉;贾春玲;袁洁;
广东省农业科学院兽医研究所广东省兽医公共卫生实验室;
Author(s):
WEI Wen-kangLV Dian-hongWEN Xiao-huiLUO Sheng-junHUANG ZhongZHOU Xiu-rongJIA Chun-lingYUAN Jie
Institute of Veterinary Medicine,Guangdong Academy of Agricultural Sciences,Guangdong Key Laboratory of Veterinary Public Health,Guangzhou 510640,China
关键词:
食源性动物组织猪瘟病毒猪蓝耳病病毒双重双色荧光定量PCR
Keywords:
food-oriented animal tissues classical swine fever virus porcine reproductive and respiratory virus duplex real-time RT-PCR based on dual labeled fluorescent probes Detection method
摘要:
为建立同时检检测食源性动物组织中猪瘟病毒(CSFV)和猪蓝耳病病毒(PRRSV)的双色荧光定量RT-PCR方法。本研究根据SCFV和PRRSV基因序列设计特异性的引物和不同荧光标记的TaqMan荧光探针,通过优化反应的体系和扩增条件,建立了能够检测食源性动物组织中SCFV和PRRSV的双重双色荧光定量PCR的方法。其检测下限为1×102拷贝/μL,而且与其他一些猪病病毒无交叉反应,具有良好的特异性。该方法重复性和稳定性试验表明其组内和组间的变异系数最高分别为4.2和4.5。对比试验表明,该方法对CSFV和PRRSV检验的敏感性为常规RT-PCR方法的200倍;该方法的建立为食源性动物组织中CSFV和PRRSV提供了有效手段,该方法特异性和敏感性较好,能够应用于临床检测。
Abstract:
To establish the method of real-time PCR for detection of classical swine fever virus(CSFV) and porcine reproductive and respiratory syndrome virus(PRRSV) in food-oriented animal tissues,the specific primers and TaqMan fluorescent probes of CSFV and PRRSV were designed and synthesized,and the amplification systems were optimized.The duplex real-time RT-PCR was established based on dual labeled FAM or VIC fluorescent probes.The method established in this study showed better amplification efficiency in detecting CSFV and PRRSV with detection limit of 102 copies,which was 200 times more sensitive than the normal PCR methods and no any reaction with other related swine viruses.The intro-and inter reproducibility were 4.2% and 4.5%,respectively.Tested on 200 clinical samples by this method showed a complete coincidence with other detection methods.This specificity and sensitivity method could be used for detection of C SFV and PRRSV in food-oriented animal tissues and clinical samples

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备注/Memo

备注/Memo:
基金:广东省科技计划项目农业攻关(2009B020307007);;广州市科技计划项目重大科技专项(2009A1-E041-1);;广东省科技计划项目促进服务业发展专项计划(2010A040207006)
更新日期/Last Update: 2011-09-29