摘要
建立一种利用环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)快速检测沙眼衣原体的方法。针对沙眼衣原体gyrA基因设计LAMP特异引物,并加入一条环引物LB提高反应效率,对反应的特异性、灵敏度和重复性进行评价,同时用沙眼衣原体阳性的临床样本进行LAMP检测验证。实时荧光LAMP结果表明,建立的LAMP方法特异性好,不与10种常见的病原微生物以及生殖道感染相关细菌发生非特异反应,灵敏度可以达到4.13×101 copies/μL;20次重复试验的Cq值变异系数为7.23%,反应稳定性良好;用LAMP方法对6份临床阳性样本进行检测,阳性符合率为100%。因此,该研究建立的沙眼衣原体LAMP检测方法快速、灵敏、稳定性好,能为感染筛查和早期诊断提供参考。
A method was developed for the rapid detection of Chlamydia trachomatis by using loop-mediated isothermal amplification(LAMP).The LAMP specific primers were designed for the gyrA gene of C.trachomatis,and a loop primer LB was added to improve the reaction efficiency.The specificity,sensitivity and repeatability of the reaction were evaluated,followed by the testing on the clinical positive samples of C.trachomatis by LAMP.Real-time fluorescent LAMP results showed that the LAMP method established had good specificity,without non-specific reaction with 10 common pathogenic bacteria associated with reproductive tract infections,and itssensitivity could reach 4.13×101 copies/μL.A total of 20 repeatingtests were performed,and the variation coefficient of Cq value was 7.23%,indicating that the stability of the LAMP methodwas satisfactory.Six clinically positive samples were used for validation of LAMP developed,and the positive coincidence rate was 100%.Accordingly the LAMP method established for detecting C.trachomatis is rapid,sensitive and stable,and it can provide a reference for screening and early diagnosis of C.trachomatis causing infection.
作者
李若琳
曹月
梁宇晨
李梦雪
朱婷
鞠丹
张彪
谢轶
许文明
孙群
LI Ruolin;CAO Yue;LIANG Yuchen;LI Mengxue;ZHU Ting;JU Dan;ZHANG Biao;XIE Yi;XU Wenming;SUN Qun(Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education,College of Life Sciences,Sichuan University,Chengdu 610064,China;Tianjin Chasesun Pharmaceutical Co.,Ltd.,Tianjin 301700,China;Chengdu institute for Food and Drug Control,Chengdu 610045,China;Department of Laboratory Medicine,West China Hospital,Sichuan University,Chengdu 610041,China;Joint Laboratory for Reproductive Medicine,SCUCUHK,West China Second University Hospital,Sichuan University,Chengdu 610041,China)
出处
《中国测试》
CAS
北大核心
2020年第4期65-69,共5页
China Measurement & Test
基金
国家重点研发计划(项目编号:SQ2019YFE010495)
成都市技术创新研发项目(2020-YF05-00306-SN)。
关键词
环介导等温扩增技术
沙眼衣原体
GYRA基因
快速检测
loop-mediated isothermal amplification technique
Chlamydia trachomatis
gyrA gene
rapid detection
