等温核酸扩增技术在食源性致病菌检测中的研究进展

等温扩增技术是近年来发展迅速的一种新型核酸扩增技术, 其反应过程始终维持在恒定的温度下, 降低了对精密仪器和检测场地的要求, 并大幅度缩短了检测时间, 更能满足准确、快速、灵敏、便携的日常检测需求。目前, 食源性致病菌仍是影响食品安全的主要因素之一, 等温核酸扩增技术已成功应用于部分食源性致病菌的检测中, 具有广阔的应用前景, 有望成为食源性致病菌快速检测的新方法。本文综述了环介导核酸等温扩增、滚环扩增、跨越式滚环扩增、重组酶聚合酶扩增、等温多自配引发扩增、单引物等温扩增、依赖解旋酶的等温扩增、链置换扩增、交叉引物扩增9种等温核酸扩增技术的扩增原理和优缺点, 及其在食源性致病菌检测中的应用研究进展, 提出了样品前处理、引物设计、结果判读、检测灵敏度、检测通量方面存在的共性问题, 并给出相应解决措施, 以期为等温核酸扩增技术在食源性致病菌快速检测中的实际应用和相关标准的制定提供研究思路。

Research progress of isothermal nucleic acid amplification techniques in the detection of foodborne pathogens

Isothermal amplification technology is a new type of nucleic acid amplification technology that has developed rapidly in recent years. Its reaction process is always maintained at a constant temperature, which reduces the requirements for precision instruments and detection sites, and greatly shortens the detection time, It can better meet the daily detection requirements of accuracy, rapid, sensitivity and portability. At present, foodborne pathogens are still one of the main factors affecting food safety. Isothermal nucleic acid amplification technology has been successfully applied to the detection of some foodborne pathogens, which has broad application prospects and is expected to become a new method for rapid detection of foodborne pathogens. This paper reviewed the amplification principles, advantages and disadvantages of the 9 kinds of isothermal nucleic acid amplification techniques, including loop-mediated isothermal amplification, rolling circle amplification, saltatory rolling circle amplification, recombinase polymerase amplification, isothermal multiple self-matching-initiated amplification, single primer isothermal amplification, helicase dependent amplification, strand displacement amplification, crossing priming amplification, and the research progress of the application in detection of foodborne pathogens. Besides, this paper proposed some drawbacks shared in sample pretreatment, primer design, result interpretation, detection sensitivity and detection flux, and given the corresponding solutions, in order to provide research ideas for the practical application of isothermal nucleic acid amplification technology in rapid detection of foodborne pathogens and the formulation of relevant standards.