多重PCR快速检测婴幼儿奶粉中的病原菌

Multiplex PCR rapid detection for pathogenic bacteria in milk powder

为了建立同时检测婴幼儿奶粉中阪崎肠杆菌、金黄色葡萄球菌、蜡样芽孢杆菌的多重PCR方法,根据阪崎肠杆菌ompA基因、金黄色葡萄球菌nuc基因、蜡样芽孢杆菌hblA基因分别设计3对引物进行多重PCR扩增,并对反应条件进行优化。结果多重PCR扩增出长度为514、156、235bp的特异性目的条带。不增菌的情况下,多重PCR同时检测3种病原菌的灵敏度是103cfu/mL,3种病原菌在奶粉中的检出限是104cfu/g。建立的多重PCR反应准确、快速、高效,为同时检测婴幼儿奶粉中的阪崎肠杆菌、金黄色葡萄球菌、蜡样芽孢杆菌提供了新方法。

To establish a multiplex PCR method for the simultaneous detection of pathogenic bacteria such as Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus in milk powder, three pairs of oligonucleotide primers were designed according to Enterobacter sakazakii ompA gene, Staphylococcus aureus nuc gene and Bacillus cereus hblA gene. They were used to amplify the special DNA sequences by multiplex PCR and the reaction conditions were optimized. The results showed that the multiplex PCR assay amplified three specific target channels of 514 bp, 156 bp and 235 bp. Without bacterial enrichment, the sensitivities of the multiplex PCR for Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus were 103 cfu/mL, and the detection limits of the multiplex PCR for Enterobacter sakazakii, Staphylococcus aureus and Bacillus cereus in milk powder were 104 cfu/g. This multiplex PCR method was accurate, fast and effective and provided a new approach for detecting Enterobacter sakazakii, Staphylococcus aureus, and Bacillus cereus in milk powder.