以金黄色葡萄球菌clfa基因和溶血性链球菌sdy基因为靶基因设计引物.通过对单个基因PCR和多重基因PCR扩增进行特异性、敏感性试验以及优化反应体系,建立了快速检测溶血性链球菌和金黄色葡萄球菌的双重PCR方法.在郑州市不同地区随机抽检126份生鲜猪肉样品,分别进行了双重PCR检测和常规微生物学检验.结果表明,建立的双重PCR方法特异性好,抗干扰能力强,灵敏度可达到102 ng·L-1.在126个样品中检测出溶血性链球菌的样品数为7份,检出率为5.55％;检出金黄色葡萄球菌阳性的样品数为8份,检出率为6.35％.%Primersfor clfa gene of Staphylococcus aureus and sdy gene of Streptococcus hemolyticus were specifically designed.Through reaction optimization and specificity and sensitivity tests,singular and duplex PCR methods were set up,and were then applied to detection of the target bacteria.126 raw pork samples collected from different places of Zhengzhou city,were analyzed by the PCR method,and conventional microbiological examination was also taken as a control.The results show that the duplex PCR protocol turned out to be specific,effective with a sensitivity of 102 ng· L-1.The examination of the raw pork samples showed that 7 out of 126 were detected as Streptococcus hemolyticus positive (5.55％),whilst Staphylococcus aureus was detected among 8 samples,showing a detection rate of 6.35％.
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