环介导等温扩增技术在快速检测炭疽芽胞杆菌中的应用

Loop-mediated isothermal amplification technology in the rapid detection of Bacillus anthracis

本文旨在建立适合国境口岸现场应用的生物恐怖防控快速检测方法,从而保障口岸安全。针对生物恐怖炭疽芽胞杆菌,选择目标菌种特异性基因片段,设计引物,运用环介导等温扩增(LAMP)技术建立一套简便、高效的检测方法,并模拟生物恐怖炭疽芽胞杆菌可能存在的基质条件,评价LAMP技术在快速筛查中的适用性。结果显示,LAMP技术排查生物恐怖炭疽芽胞杆菌简便、快速、特异,检测灵敏度为102～103CFU/ml;且能有效检出在偏酸、偏碱及黏稠基质中的炭疽芽胞杆菌。而高盐环境对该反应影响较大,有必要采用能有效去除盐分的核酸抽提方法。

The present paper aims to establish a rapid detection method for Bacillus anthracis. The primes were designed according to Bacillus anthracis strain-specific gene fragment, and loop-mediated isothermal amplification （LAMP） was used to establish the detection method. The results showed that LAMP can effectively identify the specific target bacteria with sensitivity of 102^-103^ CFU/ml. It is suggested that LAMP is simple and fast in detection of bioterrorism bacteria such as Bacillus anthracis in acidic, alkaline and viscous media. High-salt environment influences LAMP results, so it is necessary to effectively remove salt out of nucleic acid before application of LAMP.