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[目的]建立单增李斯特菌(Listeria monocytogenes,L.m)hly基因、金黄色葡萄球菌(Staphyl ococcus aureus,S.a)nuc基因、蜡样芽孢杆菌(Bacillus cereus,B.c)16S-23SITS基因的多重PCR产物的毛细管电泳-激光诱导荧光检测方法(CE)。[方法]提取细菌DNA后,分别设计扩增单增李斯特菌(Listeria monocytogenes,Lm)hly基因、金黄色葡萄球菌(Staphyl ococcus aureus,S.a)nuc基因、蜡样芽孢杆菌(Bacillus cereus)16S-23SITS基因的引物,进行三重PCR扩增,采用无胶筛分毛细管电泳作为分离手段,以激光诱导荧光检测PCR扩增产物。[结果]对多重PCR扩增产物的条件进行了优化,20min内即可完成3种致病菌的同时检测。迁移时间的日内相对标准偏差为0.92%～1.58%。[结论]所建立的方法可以同时快速检测3种食源性致病菌并应用食品样品的检测。与单一PCR扩增和传统的凝胶电泳相比,快速简便,灵敏可靠,为食源性致病菌检测提供可靠的快速检测方法,为应对突发性的公共卫生事件提供检测手段,对保障食品安全具有一定的实际意义。
[Objective] The research aimed to establish a multiplex PCR product of Listeria monocytogenes (Lm) hly gene, Staphyloccus aureus (Sa) nuc gene and Bacillus cereus (Bc) 16S-23SITS gene Capillary Electrophoresis - Laser Induced Fluorescence Detection (CE). [Method] After the bacterial DNA was extracted, the hly gene of Listeria monocytogenes (Lm), the nuc gene of Staphyloccus aureus (Sa), the gene encoding Bacillus cereus 16S- 23SITS gene primers for triplex PCR amplification, the use of non-gel separation capillary electrophoresis as a means of separation, laser-induced fluorescence detection of PCR amplification products. [Result] The conditions of multiple PCR amplification products were optimized, and the simultaneous detection of three kinds of pathogenic bacteria could be completed in 20 minutes. The relative standard deviations for the migration time ranged from 0.92% to 1.58%. [Conclusion] The established method can simultaneously detect three kinds of food-borne pathogens and apply the detection of food samples simultaneously. Compared with single PCR amplification and traditional gel electrophoresis, the method is rapid, simple, sensitive and reliable, and provides reliable and rapid detection method for foodborne pathogenic bacteria detection. In order to respond to unexpected public health events, Food safety has some practical significance.