| 马鼻肺炎病毒双重PCR分型检测方法的建立 |
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| 引用本文： | 田志革,侯玉杰,郭巍,曲娟娟,相文华.马鼻肺炎病毒双重PCR分型检测方法的建立[J].中国兽医科学,2011(9). |
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| 作者姓名： | 田志革 侯玉杰 郭巍 曲娟娟 相文华 |
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| 作者单位： | 东北农业大学生命科学学院;中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室;泰州康正生物技术有限公司; |
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| 基金项目： | 东北农业大学科学研究基金项目(200603) |
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| 摘 要： | 根据GenBank中登录的马鼻肺炎病毒1、4型(EHV1、EHV4)糖蛋白B(gB)基因特异保守序列(登录号分别为AY665713、AF030027.1),各设计1对引物,建立了同时分型检测EHV1、EHV4的双重PCR方法;确定其最佳工作条件,并对该方法进行了特异性和敏感性试验。结果显示,在EHV1、EHV4中分别扩增出265bp和537bp的特异性目的条带,而马流感病毒cDNA、马动脉炎病毒cDNA、马乙型脑炎病毒cDNA及马传染性贫血弱毒疫苗株cDNA均未扩增出任何条带。EHV1的DNA最低检出限为2.1pg,EHV4的最低检出限为15pg。表明本试验建立的方法具备很高的应用价值。
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| 关 键 词： | 马鼻肺炎病毒1、4型 双重聚合酶链反应 分型检测 |
Establishment of a duplex PCR assay for typing and detection of equine herpesviruses type 1 and 4 |
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| TIAN Zhi-ge,HOU Yu-jie,GUO Wei,QU Juan-juan,XIANG Wen-hua.Establishment of a duplex PCR assay for typing and detection of equine herpesviruses type 1 and 4[J].Veterinary Science in China,2011(9). |
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| Authors: | TIAN Zhi-ge HOU Yu-jie GUO Wei QU Juan-juan XIANG Wen-hua |
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| Institution: | TIAN Zhi-ge1,3,HOU Yu-jie2,GUO Wei3,QU Juan-juan1,XIANG Wen-hua3(1.College of Life Science,Northeast Agricultural University,Harbin 150030,China,2.Kang-zheng Bio-technology Co.Ltd.,Taizhou 225300,3.State Key Laboratory of Veterinary Biotechnology/Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China) |
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| Abstract: | According to the conserved sequences of glycoprotein B(gB) gene of equine herpesviruses(EHVs) type 1 and 4 in GenBank,a set of primers was designed to establish a duplex PCR method for detecting the two kinds of viruses.And the optimal conditions,specificity and sensitivity test were determined.In result,the PCR products for EHV1 and EHV4 were 265 bp(EHV1) and 537 bp(EHV4) with detection limit of 2.1 pg and 15 pg of DNA copies,respectively,while no product was found from closely realted virus,including equi... |
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| Keywords: | equine herpesviruses type 1 and 4 duplex PCR detection for distinction |
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