肉中两种致病菌的双重PCR检测方法的建立

为了建立肉中单增李斯特菌和金黄色葡萄球菌的双重PCR检测方法,以金黄色葡萄球菌nuc基因和单增李斯特菌hly基因为靶基因设计引物,通过优化退火温度和PCR反应体系,建立这两种菌的双重PCR检测方法,并分析其反应特异性和敏感性。进一步将该方法应用到人工模拟肉样中,并分析其敏感性。结果表明,建立的单增李斯特菌和金黄色葡萄球菌双重PCR检测方法应用于人工模拟肉样中具有较高敏感性,其中金黄色葡萄球菌和单增李斯特菌最低检测限分别为102和103 cfu.mL-1。该方法的建立为食品中多种致病菌的同时检测奠定了基础。

Establishment of duplex PCR for detection of two bacterial pathogens in meat

To detect Listeria monocytogenes and Staphylococcus aureus in meat at the same time,a duplex PCR system by was established selecting the nuc(Staphylococcus aureus) and hly(Listeria monocytogenes) genes as the target genes,which was developed by optimization of the annealing temperature and PCR system.Along with this,the specificity and sensitivity of the method were monitored.Further,the duplex PCR method was applied to detect Listeria monocytogenes and Staphylococcus aureus in artificially inoculated meat samples.The results showed that the sensitivity values of the duplex PCR mothed were around 102 cfu·mL-1 for Staphylococcus aureus and 103 cfu·mL-1 for Listeria monocyto-genes in artificially inoculated meat samples,which would show the good potential for detection of many species of bacteria in foods simultaneously.