猪繁殖与呼吸综合征病毒荧光定量RT-PCR和RT-LAMP快速检测方法的建立

中国畜牧兽医

猪繁殖与呼吸综合征病毒荧光定量RT-PCR和RT-LAMP快速检测方法的建立

周康平,郑虎，王美玉，韩振伟，王业富

(武汉大学生命科学学院，湖北武汉 430072）

出版日期:2014-01-20 发布日期:2014-02-27
通讯作者: 王业富（1962—），教授。E-mail：wangyefu@whu.edu.cn
作者简介:周康平（1989—），男，湖北人，博士生，研究方向：临床诊断和病毒机理。
基金资助:
高致病性动物疫病衍生的分子机理及预警系统（201110036-03）。

Development of Real-time RT-PCR and RT-LAMP for Rapid Detection of Porcine Reproductive and Respiratory Syndrome Virus

ZHOU Kang-ping,ZHENG Hu,WANG Mei-yu,HAN Zhen-wei,WANG Ye-fu

(College of Life Sciences,Wuhan University,Wuhan 430072,China)

Online:2014-01-20 Published:2014-02-27

摘要/Abstract

摘要： 猪繁殖与呼吸综合征病毒（porcine reproductive and respiratory syndrome virus，PRRSV）是世界各地重要猪病毒性病原之一，每年给养猪业造成重大损失。本试验建立了应用TaqMan-LNA的荧光定量RT-PCR和逆转录—环介导等温扩增(RT-LAMP)2种PRRSV检测方法。结果表明荧光定量RT-PCR和RT-LAMP检测下限分别为10²和10¹ 拷贝数/μL，都具有良好的特异性。临床样本检测结果同样表明2种方法都具有良好的特异性和灵敏度。本试验所建立的TaqMan-LNA荧光定量RT-PCR和RT-LAMP可有效的应用于PRRSV的检测。

关键词: 猪繁殖与呼吸综合征病毒；荧光定量RT-PCR；逆转录— 环节导等温扩增

Abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) were one of the worldwide causative agent for the disease of pigs and caused heavy losses every year.In this study, a TaqMan-LNA based Real-time RT-PCR assay and a reverse transcription loop-mediated isothermal amplification （RT-LAMP） assay were established.The detection limits were 10²and 10¹ copies/μL for Real-time RT-PCR and RT-LAMP assay， respectively. Two methods both showed good specificity.Futhermore both methods were analysed with clinical samples, and they exhibited high sensitivity and specificity.The Real-time RT-PCR and RT-LAMP assays would provide useful tools for the detection of PRRSV.

Key words: porcine reproductive and respiratory syndrome virus; Real-time RT-PCR; reverse transcription loop-mediated isothermal amplication

周康平,郑虎，王美玉，韩振伟，王业富. 猪繁殖与呼吸综合征病毒荧光定量RT-PCR和RT-LAMP快速检测方法的建立[J]. 中国畜牧兽医.

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