环介导等温扩增技术检测鼠疫耶尔森菌的敏感性和特异性研究

谢辉1,祁芝珍1,杨晓艳1,袁静2,李翔1,赵海红1

微生物与感染 ›› 2021, Vol. 16 ›› Issue (3) : 176-182.

PDF(3870 KB)
欢迎访问《微生物与感染》官方网站,今天是
PDF(3870 KB)
微生物与感染 ›› 2021, Vol. 16 ›› Issue (3) : 176-182. DOI: 10.3969/j.issn.1673-6184.2021.03.004
论著

环介导等温扩增技术检测鼠疫耶尔森菌的敏感性和特异性研究

  • 谢辉1,祁芝珍1,杨晓艳1,袁静2,李翔1,赵海红1
作者信息 +

Sensitivity and specificity of loop-mediated isothermal amplification in detection of Yersinia pestis

  • XIE Hui1, QI Zhizhen1, YANG Xiaoyan1, YUAN Jing2, LI Xiang1, ZHAO Haihong1
Author information +
文章历史 +

摘要

为观察环介导等温扩增(loop-mediated isothermal amplification,LAMP)技术能否适用于我国不同疫源地鼠疫耶尔森菌所有基因组型的检测,本研究建立了一种基于3a靶序列设计特异性引物快速检测鼠疫耶尔森菌的LAMP方法。选择分离自我国11个鼠疫自然疫源地的65株野生代表性鼠疫耶尔森菌株,同时以与其近源的假结核耶尔森菌、小肠结肠炎耶尔森菌及非近源的大肠埃希菌为对照菌株,观察其敏感性和特异性。结果显示,该LAMP方法与其他3种鼠疫耶尔森菌常规检测方法的结果一致。65株鼠疫耶尔森菌检测均为阳性,最低检出浓度为20个菌/μL;假结核耶尔森菌、小肠结肠炎耶尔森菌及大肠埃希菌标准菌株反应均为阴性。结果提示,本研究建立的LAMP技术对检测我国各疫源地鼠疫耶尔森菌具有高度特异性,且快速简便,无需特殊仪器,可肉眼判读结果,适用于野外现场鼠疫监测中的快速检测,在偏远地区疫情防控中具有实用价值。

Abstract

The present paper aims to study the sensitivity and specificity of loop-mediated isothermal amplification (LAMP) in detection of Yersinia pestis (Y. pestis) from all natural plague foci of China. The LAMP was established based on the specific primers targeting 3a gene of Y. pestis. Sixty-five Y. pestis strains were isolated from 11 natural plague foci of China, and Y. pseudotuberculosis, Y. enterocolitica and Escherichia coli (E. coli) were set as control strains. The results of LAMP detection were consistent with three standard diagnostic methods of plague (reverse indirect haemagglutination test, colloidal gold immunochromatography assay and conventional polymerase chain reaction method). The 65 Y. pestis strains were all positive, while Y. pseudotuberculosis, Y. enterocolitica and E. coli were negative, suggesting that the established LAMP method was highly specific for Y. pestis. It has the advantages of fast, simple, low requirements on equipment, with a potential application in field monitoring and detection in remote areas for plague prevention and control.

关键词

环介导等温扩增 / 快速检测 / 鼠疫耶尔森菌 / 鼠疫自然疫源地

Key words

Loop-mediated isothermal amplification / Rapid test / Yersinia pestis / Plague natural focus

引用本文

导出引用
谢辉1,祁芝珍1,杨晓艳1,袁静2,李翔1,赵海红1. 环介导等温扩增技术检测鼠疫耶尔森菌的敏感性和特异性研究[J]. 微生物与感染. 2021, 16(3): 176-182 https://doi.org/10.3969/j.issn.1673-6184.2021.03.004
XIE Hui1, QI Zhizhen1, YANG Xiaoyan1, YUAN Jing2, LI Xiang1, ZHAO Haihong1. Sensitivity and specificity of loop-mediated isothermal amplification in detection of Yersinia pestis[J]. Journal of Microbes and Infections. 2021, 16(3): 176-182 https://doi.org/10.3969/j.issn.1673-6184.2021.03.004
中图分类号: R446.5   

基金

国家卫生健康委员会鼠疫防治研究重点实验室(青海省地方病预防控制所)(2019PT310004),青海省鼠疫防控及研究重点实验室项目(2021-ZJ-Y15)

PDF(3870 KB)

Accesses

Citation

Detail

段落导航
相关文章

/