基于LAMP的新试剂和仪器的效果评估及在蚊媒病毒检测中的应用  

Development and Evaluation of New Reagents and Isothermal Amplification Equipment Based on LAMP and Application in Detection of Mosquito-Borne Viruses

作　　者：赵娜 赵望 刘淇 夏菡[1] 袁志明[1] ZHAO Na;ZHAO Wang;LIU Qi;XIA Han;YUAN Zhi-Ming(Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071,China;Shanghai Cohere Electronics Technology Co., Ltd, Shanghai 201611,China;Department of Environmental Science & Engineering, Fudan University, Shanghai 200433,China)

机构地区：[1]中国科学院武汉病毒研究所,湖北武汉430071 [2]上海科源电子科技有限公司,上海201611 [3]复旦大学环境科学与工程系,上海200433

出　　处：《生物技术通讯》2017年第6期838-842,共5页Letters in Biotechnology

基　　金：科技基础性工作专项(2013FY113500);中国科学院重点部署项目(ZDRW-ZS-2016-4)

摘　　要：目的:针对常见蚊媒病毒筛选引物,运用环介导等温扩增(LAMP)技术建立便捷、快速、高通量的检测方法;并以实验室储存病毒资源,评价LAMP研发试剂和仪器在快速筛查中的适用性。方法:环介导等温扩增。结果:筛选出7种蚊媒病毒高特异性引物,检测用研发试剂为单管固定化反应管,反应只需加入模板,使用配套仪器,检测总耗时不超过45 min,检测灵敏度为10^(-1)~10~2拷贝。结论:研发仪器一键式操作、便携轻便,可实时监测;研发试剂单管储存固定化反应管,极大程度地降低了污染,简单快捷,扩增稳定重复性好且成本较商业化试剂低;在现场实现蚊媒病毒的简便、快速、低成本、实时高通量检测。Objective:To develop a rapid,convenient,and high throughput detection assay of common mosquitoborne viruses based on loop-mediated isothermal amplification(LAMP),and evaluate the applicability of new reagents and equipment in rapid detection.Methods:Loop-mediated isothermal amplification.Results:Seven primer sets of mosquito-borne viruses with high specificity were screened.When combine used the new reagents(a type of immobilized reaction tubes)and equipment,the detection for7mosquito-borne viurses can be finished within45min with the limitation around10-1~102copies.Conclusion:The developed equipment is one-touch operation,mobile and light while real-time monitor.Immobilized reaction tubes needing templates only can reduce the contamination,and the amplification is stable and repeatable and much cheaper than commercial reagents which implement a rapid,convenient,low-cost and high throughput detection of common mosquito-borne viruses.

关 键 词：环介导等温扩增 蚊媒病毒 固定化反应管 恒温扩增荧光读数仪 现场快速检测 

分 类 号：Q78[生物学—分子生物学]