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目的:探索建立一种能应用于单增李斯特菌检测与鉴定的LAMP方法,以适合在基层实验室开展应用。方法:针对单增李斯特菌invA基因靶序列,设计5条特异性LAMP引物,建立LAMP反应条件与体系。结果:整个检测过程仅需1.5 h,可通过肉眼目测或电泳检测判断结果,对5株单增李斯特菌和英诺克李斯特菌等共23株其它实验株的invA基因进行检测,结果显示该方法有良好的特异性;检测限试验显示敏感性稍差但不影响对单增李斯特菌分离疑似菌株鉴定及疑似食品污染物的批量检测。结论:本研究建立的LAMP方法能够快速、特异地检测单增李斯特菌;简单经济,不需要其他辅助仪器,结果即可直观判断;适合基层实验室、应急检测或现场监测等使用,具有较高的推广价值。
Objective: To establish a LAMP method that can be applied to the detection and identification of Listeria monocytogenes, which is suitable for the application in grassroots laboratories. Methods: According to the target sequence of invA gene of Listeria monocytogenes, five specific LAMP primers were designed and LAMP reaction conditions and system were established. Results: The whole detection process was only 1.5 h. The results of visual inspection or electrophoresis were used to detect the invA gene of 5 strains of Listeria monocytogenes and Listeria monocytogenes. The results showed that The method has good specificity; the limit of detection test shows less sensitivity but does not affect the identification of suspicious strains isolated from Listeria monocytogenes and the batch detection of suspected food contaminants. Conclusion: The LAMP method established in this study can detect Listeria monocytogenes rapidly and specifically. It is simple and economical, does not need other auxiliary instruments, and can be judged intuitively. It is suitable for primary laboratory, emergency detection or on-site monitoring, High promotion value.