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[耐甲氧西林金黄色葡萄球菌——mecA及其调控基因的检测]

[MRSA--detection of mecA and its regulatory genes].

作者信息

Kagawa S, Yamashita K, Matsuoka A

机构信息

Clinical Laboratory, Hyogo College of Medicine, Nishinomiya.

出版信息

Rinsho Byori. 1993 Nov;41(11):1223-31.

PMID:8283799
Abstract

Methicillin resistance in S.aureus and S.epidermidis strains is primarily due to production of a new penicillin-binding protein PBP2' with extremely low binding affinity to most beta-lactam antibiotics. The structural gene for PBP2', mecA, is detectable in clinical specimens by using the polymerase chain reaction (PCR). Amplified target DNA of 630bp can be resolved on ethidium bromide-stained gels, and hybridized with a probe conjugated to alkaline phosphatase. Survey for the mecA gene in 304 staphylococci revealed a good correlation between the presence of mecA and cultivation on agar plates with 4 micrograms/ml of oxacillin, although 3% of sensitive S. aureus strains had the mecA gene. On the other hand, analysis of the regulatory genes (orf 1 and 2) of methicillin resistance was performed on methicillin-resistant S.aureus strains N315 and MR108, demonstrating that the genome of MR108 lacks orf 2 which encodes the repressor protein (Hiramatsu et al., see Ref.5). The regulatory genes of mecA were surveyed for 192 staphylococci by using PCR and allele-specific oligonucleotide probes: 76% of resistant S. aureus strains and 48% of resistant S. epidermidis strains possessed orf 1 corresponding to MR108 (constitutive-type strain), while the remainder of the resistant strains and two strains of sensitive S. epidermidis had two orfs of N315 (inducible-type strain). Furthermore, it appeared that mutation of the femA gene might not be an additional factor for expression of methicillin resistance. These observations suggest that mecA and its regulatory genes should be examined to understand how the genetic background contributed to the phenotypic expression of methicillin resistance in clinical strains.

摘要

金黄色葡萄球菌和表皮葡萄球菌菌株中的耐甲氧西林特性主要归因于一种新的青霉素结合蛋白PBP2'的产生,该蛋白对大多数β-内酰胺类抗生素的结合亲和力极低。通过聚合酶链反应(PCR)可在临床标本中检测到PBP2'的结构基因mecA。扩增得到的630bp靶DNA可在溴化乙锭染色的凝胶上分辨出来,并与碱性磷酸酶偶联的探针杂交。对304株葡萄球菌进行的mecA基因检测显示,mecA的存在与在含4微克/毫升苯唑西林的琼脂平板上培养之间存在良好的相关性,尽管3%的敏感金黄色葡萄球菌菌株含有mecA基因。另一方面,对耐甲氧西林金黄色葡萄球菌菌株N315和MR108进行了耐甲氧西林调控基因(orf 1和2)的分析,结果表明MR108的基因组缺乏编码阻遏蛋白的orf 2(平松等人,见参考文献5)。通过PCR和等位基因特异性寡核苷酸探针,对192株葡萄球菌的mecA调控基因进行了检测:76%的耐甲氧西林金黄色葡萄球菌菌株和48%的耐甲氧西林表皮葡萄球菌菌株拥有与MR108相对应的orf 1(组成型菌株),而其余的耐甲氧西林菌株和两株敏感表皮葡萄球菌菌株具有N315的两个orf(诱导型菌株)。此外,femA基因的突变似乎不是耐甲氧西林表达的额外因素。这些观察结果表明,应该检测mecA及其调控基因,以了解遗传背景如何影响临床菌株中耐甲氧西林表型的表达。

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