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多重聚合酶链反应直接从血培养瓶中检测 mecA 基因和鉴定金黄色葡萄球菌。

Detection of the mecA gene and identification of Staphylococcus directly from blood culture bottles by multiplex polymerase chain reaction.

机构信息

UNESP - Univ Estadual Paulista, Instituto de Biociências de Botucatu, Departamento de Microbiologia e Imunologia, Botucatu, SP, Brazil; UNESP - Univ Estadual Paulista, Faculdade de Medicina de Botucatu, Hospital Universitário, Departamento de Doenças Tropicais, Botucatu, SP, Brazil.

UNESP - Univ Estadual Paulista, Instituto de Biociências de Botucatu, Departamento de Microbiologia e Imunologia, Botucatu, SP, Brazil.

出版信息

Braz J Infect Dis. 2018 Mar-Apr;22(2):99-105. doi: 10.1016/j.bjid.2018.02.006. Epub 2018 Mar 13.

Abstract

INTRODUCTION

Staphylococcus spp. - both S. aureus, including methicillin-resistant strains (MRSA) and coagulase negative staphylococci (CoNS) - are relevant agents of healthcare-associated infections. Therefore, the rapid recognition of MRSA and methicillin-resistant CoNS from blood stream infections is critically important for patient management. It is worth noting that inappropriate empiric therapy has been associated with higher in-hospital mortality.

MATERIAL AND METHODS

In this study we evaluated a multiplex polymerase chain reaction (multiplex PCR) standardized to detect Staphylococcus spp., S. aureus, and mecA gene-encoded oxacillin resistance directly from blood culture bottles. A total of 371 blood cultures with Gram-positive microorganisms confirmed by Gram-stain were analyzed. Results from multiplex PCR were compared to phenotypic characterization of isolates.

RESULTS

Staphylococcus aureus was detected in 85 (23.0%) blood cultures and CoNS in 286 (77.0%). There was 100% agreement between phenotypic and multiplex PCR identification. Forty-three (50.6%) of the 85 S. aureus carried the mecA gene and among the 286 CoNS, 225 (78.7%) were positive for the mecA gene.

CONCLUSIONS

The multiplex PCR assay developed here was found to be sensitive, specific, rapid, and showed good agreement with the phenotypic results besides being less expensive. This PCR method could be used in clinical laboratories for rapid identification and initiation of specific and effective treatment, reducing patient mortality and morbidity. Furthermore, this method may reduce misuse of antimicrobial classes that are more expensive and toxic, thus contributing to the selection of antibiotic-resistant Staphylococcus spp.

摘要

简介

金黄色葡萄球菌(包括耐甲氧西林金黄色葡萄球菌(MRSA)和凝固酶阴性葡萄球菌(CoNS))是与医疗保健相关感染相关的重要病原体。因此,快速识别血流感染中的 MRSA 和耐甲氧西林凝固酶阴性葡萄球菌对于患者管理至关重要。值得注意的是,经验性治疗不当与住院死亡率升高有关。

材料和方法

在这项研究中,我们评估了一种多重聚合酶链反应(multiplex PCR),该方法经过标准化,可直接从血培养瓶中检测金黄色葡萄球菌、金黄色葡萄球菌和 mecA 基因编码的耐甲氧西林。总共分析了 371 个革兰氏阳性微生物的血培养物,这些微生物通过革兰氏染色得到确认。将多重 PCR 的结果与分离株的表型特征进行比较。

结果

在 85 个血培养物中检测到金黄色葡萄球菌,在 286 个血培养物中检测到凝固酶阴性葡萄球菌。表型和多重 PCR 鉴定之间存在 100%的一致性。85 株金黄色葡萄球菌中有 43 株(50.6%)携带 mecA 基因,286 株凝固酶阴性葡萄球菌中有 225 株(78.7%) mecA 基因阳性。

结论

这里开发的多重 PCR 检测方法具有敏感性、特异性、快速性和与表型结果良好的一致性,同时价格更低。该 PCR 方法可用于临床实验室,用于快速鉴定并启动特异性和有效的治疗,降低患者的死亡率和发病率。此外,该方法可能减少对抗生素类药物的滥用,这些药物更昂贵且毒性更大,从而有助于选择耐抗生素的葡萄球菌。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/370a/9428231/08eead962934/gr1.jpg

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