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马动脉炎病毒RT-LAMP检测方法的建立

Development of an RT-LAMP assay for the detection of equine arteritis virus

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【作者】 黄素文; 于纪棉; 倪健波; 王建峰; 杨文潮; 张超; 张吉红; 相文华;

【Author】 HUANG Su-wen1,2,YU Ji-mian3,NI Jian-bo2,WANG Jian-feng2,YANG Wen-chao2, ZHANG Chao2,ZHANG Ji-hong2,XIANG Wen-hua4(1.College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China;2.Ningbo Entry-Exit Inspection and Quarantine Bureau,Ningbo 315012,China;3.Ningbo College of Health Sciences,Ningbo 315100,China; 4.Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China)

【机构】 南京农业大学动物医学院; 宁波出入境检验检疫局; 宁波天一职业技术学院; 中国农业科学院哈尔滨兽医研究所;

【摘要】 为建立一种快速、简便、特异性高的马动脉炎病毒反转录环介导等温扩增(RT-LAMP)检测方法,针对马动脉炎病毒膜基质蛋白M基因的保守序列设计特异性引物,优化反应条件,建立了马动脉炎病毒的RT-LAMP检测方法并对其特异性和灵敏性进行了检测。结果显示,建立的RT-LAMP检测方法具有良好的特异性,灵敏度为0.12pg,是普通RT-PCR的100倍,检测时间仅需1h,肉眼即可观察检测结果。表明建立的RT-LAMP检测方法可用于进出口马动脉炎病毒的检疫。

【Abstract】 A reverse transcription loop-mediated isothermal amplification(RT-LAMP) assay was established for rapid,simple and specific detection of equine arteritis virus(EAV) based on a set of primers designed according to the membrane matrix protein M gene sequence of EAV available in the GenBank.The specificity and sensitivity of the RT-LAMP were determined under the optimized amplification condition.The RT-LAMP assay was highly specific and the detection limitation was 0.12 pg,which was about 100 times higher than that of RT-PCR.It can be conducted within 1 h and the detection results were visualized.The rapid,sensitive and specific RT-LAMP assay was therefore a potential useful technique for EAV quarantine.

  • 【文献出处】 中国兽医科学 ,Chinese Veterinary Science , 编辑部邮箱 ,2011年09期
  • 【分类号】S858.21
  • 【被引频次】2
  • 【下载频次】108
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