基孔肯雅病毒LAMP检测方法的建立及其应用研究  

Establishment and application of LAMP for detecting Chikungunya virus

作　　者：李小波[1] 鲁曦[2] 幸芦琴[1] 郑夔[1] 单潇潇[2] 黄吉城[1] 石磊[2] 

机构地区：[1]广东出入境检验检疫局检验检疫技术中心卫生检疫实验室,广州510700 [2]华南理工大学轻工与食品学院,广州510640

出　　处：《中国卫生检验杂志》2012年第3期506-508,共3页Chinese Journal of Health Laboratory Technology

基　　金：国家质检总局科研项目(2010IK222);广东检验检疫局科研项目(2009GDK11)

摘　　要：目的:建立快速、灵敏、特异的基孔肯雅病毒(CHIKV)环介导等温基因扩增(LAMP)检测方法,将该方法应用于基孔肯雅热(CHIK)可疑患者的快速筛查。方法:根据CHIKV核酸序列多重比对结果设计并合成LAMP专用的扩增引物,摸索最佳反应条件,建立灵敏、特异的CHIKV核酸LAMP检测新方法;应用该方法对疑似患者血清中提取的核酸进行快速检测。结果:本方法对CHIKV样本的检测灵敏度达到27拷贝/反应,检测灵敏度高于普通PCR法,略低于实时荧光PCR法。本方法快速、灵敏,并具有较高的特异性和很好的重复性,可在两小时内对CHIK疑似病例进行准确诊断。利用本方法对入境CHIK患者及国内患者样本进行检测,均取得了良好的效果。结论:本方法适合国境口岸一线及基层卫生机构对CHIK可疑患者进行快速检测,对防止CHIK疫情向我国的传播及保障我国公共卫生安全均具有重要意义。Objective： To establish Loop-Mediated Isothermal Amplification Method for detecting CHIKV.Methods： The research prepared positive sample of CHIKV,designed and synthesized corresponding primers for LAMP to detect and find out the best condition for CHIKV.Results： The method reached 27 copies per reaction to the sensitivity of measuring CHIKV samples,which was fast and sensitive with high specifity and repetitiveness,having the capacity to examine nucleic acid of CHIKV accurately within two hours.Conclusion： The method is suitable for the detection of CHIKV at port or internal basic medical institution,which has the vital significance of preventing CHIKV spreading to our country and safeguarding the health security of our country.

关 键 词：基孔肯雅病毒 环介导等温基因扩增 

分 类 号：R446.1[医药卫生—诊断学]