三重聚合酶链反应快速诊断耐甲氧西林葡萄球菌感染

目的建立一种能快速有效地对葡萄球菌感染进行病原体诊断的方法.方法利用三重聚合酶链反应技术(Triplex PCR)同时检测葡萄球菌特异性16S rRNA 基因、金黄色葡萄球菌特异性Sa442基因和编码青霉素结合蛋白(PBP2a)的mecA基因.结果 84株葡萄球菌的16S rRNA 基因100%阳性,其中33株金黄色葡萄球菌的Sa442基因100%阳性,mecA基因阳性率为78.8%.51株凝固酶阴性葡萄球菌的Sa442基因100%阴性,mecA基因阳性率为76.5%.30株其他临床常见菌16S rRNA、Sa442、mecA基因100%阴性.结论该技术具有简便、快速、特异性强等特点,是一种非常有效的耐甲氧西林葡萄球菌感染快速诊断方法.

Triplex polymerase chain reaction for the diagnosis of methicillin-resistant staphylococcal infection

Objective To develop a rapid and effective method for the diagnosis of methicillin resistant staphylococcal infection.Methods Triplex polymerase chain reaction was developed for simultaneously detecting the 16S rRNA gene that are unique to staphylococcus,the S.aureus specific Sa442 gene and mecA gene encoding PBP2a for methicillin resistance.Results For 84 strain staphylococcus,100% showed positive for 16SrRNA gene.For 33 strain of S.aureus,100% showed positive for Sa442 gene,78.8%(26/33) positive for mecA gene,while 75.7%(25/33) MRSA was positive in antibiotic susceptibility assays.For 51 strains of coagulase negative staphylococcus(CNS),100%showed negative for Sa442 gene,76.5%(39/51) positive for mecA gene,while 80.4%(41/51) MRCNS was positive in antibiotic susceptibility assay.All 30 tested strains of non staphylococcus were negative for 16S rRNA,Sa442 gene and mecA.Conclution The results proved that triplex PCR for the diagnosis of methicillin resistant staphylococcal infection was a rapid,simple and specific method.It can be used as a efficient diagnostic method.