犬瘟热病毒RT-LAMP检测方法的建立及初步应用

›› 2012, Vol. 39 ›› Issue (10): 45-49.

犬瘟热病毒RT-LAMP检测方法的建立及初步应用

胡嘉欣^1,2, 温永俊², 霍志云^1,2, 师新川², 杨博超², 武华², 高维凡¹

1. 沈阳农业大学,辽宁沈阳 110161;
2. 中国农业科学院特产研究所,吉林长春 130122

收稿日期:2012-03-05 出版日期:2012-10-20 发布日期:2012-10-19
通讯作者: 高维凡,研究方向:免疫与微生物。E-mail:gaoweifansyau@163.com E-mail:gaoweifansyau@163.com
作者简介:胡嘉欣(1986-),男,山东人,硕士生,研究方向:免疫与微生物学。

Establishment and Preliminary Application Development of a Loop-mediated Isothermal Amplification Assay for Detection of Canine Distemper Virus

HU Jia-xin^1,2, WEN Yong-jun², HUO Zhi-yun^1,2, SHI Xin-chuan², YANG Bo-chao², WU Hua², GAO Wei-fan¹

1. Shenyang Agricultural University,Shenyang 110161,China;
2. Institute of Special Economic Animal and Plant Science, Chinese Academy of Agricultural Sciences, Changchun 130122,China

Received:2012-03-05 Online:2012-10-20 Published:2012-10-19

摘要/Abstract

摘要： 环介导等温基因扩增技术(loop-mediated isothermal amplification,LAMP)是一种特异、灵敏、快速的检测方法。为提高犬瘟热病毒(CDV)检测效率、降低检测成本,针对犬瘟热病毒N基因保守区的8个位点设计了6条引物进行RT-LAMP一步法扩增。对反应体系及条件优化,检测特异性、敏感性,扩增产物通过凝胶电泳、显色反应和浑浊度比较进行判定。试验结果表明,该方法特异性强、敏感度高,操作简便、快速、便于观察,适用于门诊和现场检测。

关键词: 犬瘟热病毒; 环介导等温基因扩增技术; N基因

Abstract: This paper developed a new method in rapid detection of canine distemper virus(CDV) based on loop-mediated isothermal amplification(LAMP)assay which overcame the difficulties found in other measures,such as efficiency and high cost. In order to improve the specificity and sensitivity, we designed 6 primers recognizing the 8 conservative sequences on N gene of CDV. The result could be easily confirmed by agarose gel electrophoresis, color reaction or turbidity comparison. In conclusion, The newly developed RT-LAMP detection method for CDV was more specific, sensitive, simple and rapid than the others, even more the proceeding reaction was visible by the naked eye, and that was also to say, it offered a simple solution in field veterinary diagnose for CDV control.

Key words: canine distemper virus(CDV); loop-mediated isothermal amplification(LAMP); N gene

中图分类号:

S852

胡嘉欣, 温永俊, 霍志云, 师新川, 杨博超, 武华, 高维凡. 犬瘟热病毒RT-LAMP检测方法的建立及初步应用[J]. , 2012, 39(10): 45-49.

HU Jia-xin, WEN Yong-jun, HUO Zhi-yun, SHI Xin-chuan, YANG Bo-chao, WU Hua, GAO Wei-fan. Establishment and Preliminary Application Development of a Loop-mediated Isothermal Amplification Assay for Detection of Canine Distemper Virus[J]. , 2012, 39(10): 45-49.

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