金黄色葡萄球菌焦磷酸测序检测及耐药研究  

Pyrophosphoric Acid Sequencing and Drug Resistance Study of Staphylococcus Aureus

作　　者：袁慕云[1] 卓锦雪[1] 许龙岩[1] 陈碧玲[1] 张旺[1] 

机构地区：[1]广东出入境检验检疫局,广东广州510623

出　　处：《检验检疫学刊》2013年第6期39-43,共5页Journal of Inspection and Quarantine

基　　金：广东出入境检验检疫局科研项目(2012GDK35)

摘　　要：[目的]探讨用norA基因特异检测金黄色葡萄球菌方法及耐药情况。[方法]根据金黄色葡萄球菌的norA基因设计扩增引物和测序引物,先用PCR特异性地扩增目的片段,再制备单链模板在测序引物引导下进行焦磷酸测序,将测序结果与GenBank中的norA基因序列比对进行鉴定;用Vitek2进行药敏试验,分析金黄色葡萄球菌耐药情况。[结果]扩增引物和测序引物表现出良好的特异性,28株金黄色葡萄球菌均扩增出大小113bp的DNA片段,焦磷酸测序结果与norA基因序列100%匹配,而阴性对照菌株均未扩增出DNA条带,焦磷酸测序结果阴性;28株金黄色葡萄球菌有14种耐药谱,青霉素、红霉素、苯唑西林和四环素的耐药率分别为71.4%、35.7%、32.1%和28.6%;6株菌株发生C-T突变,对喹诺酮类抗生素菌敏感。[结论]用norA基因可特异性的检测金黄色葡萄球菌;norA基因第354和377碱基位点的突变和喹诺酮类抗生素耐药性无关。The method of inspecting staphylococcus aureus with norA gene was discussed. To design the amplifying probe and sequencing probe according to norA gene were designed. Firstly, the gene fragment was amplified. Then the single strain probe was prepared and the pyrophosphoric acid sequencing, implemented and the seguence, compare with the one in GeneBank. Carry out the medicine sensitivity was tested and the drug resistance, analyzed Amplifying and sequencing probes both showed good specificity, and all of the 28 strains got a DNA fragment of 113bp. Pyrophosphoric acid sequencing matched the nora gene sequence with a degree of 100 percent. The negative controls showed the opposite. The 28 strains showed 14 types of drug resistance. The resistance rates of penicillin, erythrocin, oxacillin and tetracycline were 71.4%, 35.7%, 32.1% and 28.6% respectively. Six strains had C-T mutations, and were sensitive to quinolones. The norA gene could be used to inspect staphylococcus aureus. The 354 and 377 base mutations had little relation with resistance to quinolones.

关 键 词：金黄色葡萄球菌 PCR 焦磷酸测序 耐药 检测 

分 类 号：Q93-3[生物学—微生物学]