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【目的】探讨、优化基于环介导等温扩增技术(LAMP)快速检测常规食品中感染性痢疾志贺氏菌的方法。【方法】在NCBI数据库中搜索获取志贺氏菌的特异性基因高度保守区,设计3对LAMP反应引物,建立、优化该LAMP可视化检测方法,并评价其特异性、灵敏度,同时与PCR检测方法和传统检测方法对比,进行结果统计分析。【结果】5株志贺氏菌标准菌株样品均检测为阳性,11株非志贺氏菌标准菌株样品均检测为阴性,无交叉反应。最低检验限为1.6×101 CFU/反应(或1.6×101 CFU/m L),且经比较,LAMP检测灵敏度比PCR检测高出1个数量级。通过对161份实际样品和人工污染样品进行检测,LAMP检测与传统方法检测结果具有较高的一致性。【结论】LAMP具有检测过程快速简便、检测结果稳定、可靠的特点,适用于对常规食品中感染性痢疾志贺氏菌的高效、快速检测需求。
【Objective】 To explore and optimize the rapid detection of Shigella dysenteriae in conventional food based on ring-mediated isothermal amplification (LAMP). 【Method】 Three pairs of LAMP reaction primers were designed and synthesized in the NCBI database to search for the highly conserved region of specific genes of Shigella. The visualization method of LAMP was designed and optimized, and its specificity and sensitivity were evaluated. Simultaneously, Compared with the traditional detection methods, the results of statistical analysis. 【Result】 The results showed that all five standard strains of Shigella were positive and 11 of the standard strains of Shigella were negative, with no cross-reaction. The lowest limit of detection was 1.6 × 101 CFU / reaction (or 1.6 × 101 CFU / mL), and by comparison, LAMP detection sensitivity was one order of magnitude higher than PCR detection. By testing 161 real samples and artificial contaminated samples, LAMP detection has higher consistency with the traditional method. 【Conclusion】 LAMP has the advantages of fast and easy detection, stable and reliable detection results, and is suitable for the rapid and efficient detection of Shigella dysenteriae in conventional foods.