食源性致病菌等温扩增检测技术的研究进展

快速可靠的食源性致病菌检测方法是保障食品安全的关键。然而,传统培养法的检测周期过于冗长,无法适应对时间敏感度较高的现场或在线检测。为此,研究人员开发了多种以快速和高灵敏度为主要特征的等温扩增技术及相应的检测产品,为食源性致病菌的现场或在线检测提供了强有力的技术支撑。该文对环介导等温扩增、滚环扩增、链置换扩增、切口酶信号扩增、核酸外切酶Ⅲ辅助扩增5种等温扩增方法的原理及其优缺点进行了梳理和比较。在此基础上,分析了等温扩增方法在细菌活性识别、非特异性扩增、前处理效率、检测通量4个方面所遇到的共性问题,并结合作者已有工作,提出了解决这些问题的方法或策略。

Research Progress of Isothermal Amplification Techniques for Detection of Food-borne Bacteria

Rapid and reliable detection methods for food-borne pathogen are the key points for food safety control.However,traditional culture-based procedure for bacterial identification,generally taking 5 to 7 days and requiring skilled person and non-portable equipment,is incompetent for point-of-care or on-site detection.Thus,several isothermal amplification methods characterized by rapidness and thermal cycle independent were developed to fulfill some specific needs for point-of-care or on-site detection.In this review,mechanisms for isothermal amplification methods,designated as loop-mediated isothermal amplification,rolling circle amplification,strand displacement amplification,nicking enzyme signal amplification,exonuclease Ⅲ assisted amplification,were explained and compared in detail.Besides,some drawbacks shared by these methods were highlighted,and ways to overcoming those drawbacks such as adoption of propidium monoazide,novol fluorescent dye,high-throughput pretreatment equipment and microfluidic chip were provided based on the recent publications of the author s group and others.