【摘要】 目的 应用一种敏感快速的多重聚合酶链反应(PCR)，检测耐甲氧西林的金黄色葡萄球菌(MRSA)和凝固酶阴性葡萄球菌(MRCNS)。方法 临床分离的北京地区金黄色葡萄球菌123株，MRCNS122株，用溶壁素和蛋白酶K制备模板DNA，设计葡萄球菌甲氧西林耐药的决定基因，金黄色葡萄球菌独有的一个辅助基因和细菌中均有的16SrRNA基因引物，通过多重PCR技术对标本进行扩增。结果 123株金黄色葡萄球菌的femA基因100%(123/123)阳性，mecA基因阳性的占18.7%(23/123),122株MRCNS的femA100%(122/122)阴性，mecA阳性的占24.6%(30/122)。16S rRNA基因片断在多重PCR中作为内部参照避免了假阴性结果的出现。结论 建立的多重PCR技术检测MRSA和MRCNS具有敏感、快速、特异的特点，是一种可靠的实验诊断手段。

ZHENG Bo, LI Jiatai.

    Institute of Clinical Pharmacology, First Hospital, Beijing Medical University, Beijing 100034

【Abstract】 Objective To establish a sensitive and speedy multiple polymerase chainreaction (multiplex PCR) technique for methicillin resistant staphylococcus aureus andmethicillin resistant coagulase negative staphylococci detection.Methods 123 strains staphylococcus aureusand 122 strains coagulase negative staphylococci were detected for mecA, femA and 16S rRNAgenes. Template DNA of bacteria was prepared by Lysostaphine and Proteinase K. By using amultiplex PCR strategy, 392 bp and 686 bp regions of the mecA and femA genes, werecoamplified to identify susceptible (lacking mecA) and resistant (mecA⁺ )staphylococci and to differentiate staphylococcus aureus (femA⁺ ) from coagulase-negative staphylococci (laking femA). The 876 bp region of the 16S rRNA gene wasused as a PCR internal control.Results The femA gene positive rate of 123 staphylococcus aureus and 122 coagulase-negative staphylococcus was 100%(123/123) and 0%(0/122) respectively. The mecAgene positive rate of staphylococcus aureus and coagulase-negative staphylococcus was18.7% (23/123) and 24.6% (30/122).Conclusion The multiplex polymerase chain reaction is a specific, sensitiveand fast method for MRSA and MRCNS detection.

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