聚合酶链反应检测耐甲氧西林金黄色葡萄球菌的研究

STUDY ON POLYMERASE CHAIN REACTION ASSAY FOR THE DETECTION OF METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS

为对聚合酶链反应（ＰＣＲ）扩增ＭｅｃＡ基因诊断耐甲氧西林金黄色葡萄球菌（ＭＲＳＡ）的可行性进行评价。应用ＰＣＲ方法检测１２０株保留的金黄色葡萄球菌菌株，扩增结果与药敏检测结果进行比较，发现４０株对甲氧西林、苯唑西林耐药的金黄色葡萄球菌ＰＣＲ扩增结果均为阳性，而有４株菌株对甲氧西林苯唑西林敏感的金黄色葡萄球菌ＰＣＲ扩增结果阳性。其扩增产物经Ｓｏｕｔｈｅｒｎ杂交证实为ＭｅｃＡ基因特异产物，采用菌落计数的方法发现５６．７０±１２．９４个细菌即可出现阳性结果，ＰＣＲ是一快速、敏感、特异的ＭＲｓＡ诊断方法。

To evaluate the detection method of methicillin-resistant staphylococcus aureus MRSA)using polymerase chain reaction(PCR) to amplify MecA gene.120 Staphylo- coccus aureus strains were detected using PCR method. The PCR results were comparedwith that of drug susceptibility test. All 0f the strains 40 strains)resistant to methicillin and oxacillin were positive after PCR amplification.. 4 strains sensitive to methicillin and oxacillin were positive after PCR amplification。 The PCR products were proved to be specific using southern hybridization method. After 35 cycles, the positive PCR results can be gained as few as 56.70 ± 12.94 MRSA cells.It wassuggested that PCR amplification was a rapid, sensitive and specific method for the detection of MRSA。