多重PCR快速检测化妆品中三种致病菌的研究

Rapid detection of three types of bacterial pathogens in cosmetics by multiplex PCR

利用多重PCR技术建立快速检测化妆品中三种致病菌的方法。根据已报道的大肠杆菌phoA基因、铜绿假单胞菌外膜蛋白基因oprL和金黄色葡萄球菌特异性序列SmaI选择特异性引物,对人工染菌化妆品进行多重PCR检测。结果显示,三种致病菌的基因组DNA均可与各自引物特异性结合,扩增产物大小分别为622 bp、504 bp和426 bp。该方法用于人工污染的化妆品中,大肠杆菌的检出限浓度为10^3CFU/m L,铜绿假单胞菌和金黄色葡萄球菌的检出限浓度为105CFU/m L。作者建立的多重PCR方法可同时快速、特异地对化妆品中三种致病菌进行检测,在化妆品行业具有较大的应用价值。

A multiplex PCR （ m-PCR） method for rapid detection of three types of bacterial pathogens in cosmetics was developed. According to the reported, three pairs of primers were chosen for the m-PCR detection of artificially contaminated cosmetics, based on the phoA gene of Escherichia coli, the outer membrane protein （ OMP ） oprL gene of Pseudomonas aeruginosa and the SmaI specific sequence of Staphylococus aureus. The results indicated the three pairs of primers bound specifically to the genomic DNA of the three types of bacterial pathogens and amplified three fragments of 622 bp for Escherichia coli, 504 bp for Pseudomonas aeruginosa and 426 bp for Staphylococcus aureus. In the case of artificially contaminated cosmetics, the detection limits of 10^3 CFU/mL for Escherichia coli were achieved, and the detection limits of 105 CFU/mL for Pseudomonas aeruginosa were achieved as well as Staphylococus aureus. This m-PCR method could provide a simultaneous, rapid and specific detection for three types of bacterial pathogens in cosmetics, and have greater practical application values.