3种致病菌多重PCR检测体系的建立及应用

金黄葡萄球菌(Staphylococcus aureus,SA)、单核细胞增生性李斯特菌(Listeria monocytogen,LM)和蜡样芽孢杆菌(Bacillus cereus,BC)是食品中重要的致病菌,建立其多重PCR的快速检测体系,对开发食源性致病微生物快速检测试剂盒具有重要意义。根据SA的nuc基因、LM的hly基因和BC的hemolysin基因,设计合成3对特异性引物,然后进行单基因PCR反应特异性验证。在此基础上建立了3种致病菌的多重PCR检测体系,并应用于食品检测中,同时以国标法进行对比验证。结果表明,建立的多重PCR检测方法简单、快速、灵敏度高,检测灵敏度可达到1 cfu/mL,整个检测时间在16 h以内,具有较大的应用价值,可广泛应用于食品卫生检测以及临床检验等领域。

Establishment and Application of Multiplex PCR for Detection of Pathogenic Microorganism

Staphylococcus aureus(SA),Listeria monocytogen(LM) and Bacillus cereus(BC) are important food-borne pathogens in food.The establishment of the multiplex PCR detection method is very important for developing the food-borne pathogenic microorganism detection kit.Three pairs of primers were designed according to nuc gene of SA,hly gene of LM and hemolysin gene of BC,and then the PCR was performed to detect their specificities using the conventional methods as control.The results showed that the multiplex PCR method was simple,rapid and sensitive.The sensitivity achieved was as low as 1 CFU/mL and the total detection time was less than 16 hours.It could be widely applied to many fields such as food sanitation detection,clinical inspection and so on.