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通过实时聚合酶链反应(PCR)区分金黄色葡萄球菌和凝固酶阴性葡萄球菌,包括与传统微生物检测相比检测耐甲氧西林菌株。

Differentiation between Staphylococcus aureus and coagulase-negative Staphylococcus species by real-time PCR including detection of methicillin resistants in comparison to conventional microbiology testing.

作者信息

Klaschik Sven, Lehmann Lutz E, Steinhagen Folkert, Book Malte, Molitor Ernst, Hoeft Andreas, Stueber Frank

机构信息

Department of Anaesthesiology and Intensive Care Medicine, Universitätsklinikum Bonn, Bonn, Germany.

出版信息

J Clin Lab Anal. 2015 Mar;29(2):122-8. doi: 10.1002/jcla.21739. Epub 2014 May 5.

Abstract

BACKGROUND

Staphylococcus aureus has long been recognized as a major pathogen. Methicillin-resistant strains of S. aureus (MRSA) and methicillin-resistant strains of S. epidermidis (MRSE) are among the most prevalent multiresistant pathogens worldwide, frequently causing nosocomial and community-acquired infections.

METHODS

In the present pilot study, we tested a polymerase chain reaction (PCR) method to quickly differentiate Staphylococci and identify the mecA gene in a clinical setting.

RESULTS

Compared to the conventional microbiology testing the real-time PCR assay had a higher detection rate for both S. aureus and coagulase-negative Staphylococci (CoNS; 55 vs. 32 for S. aureus and 63 vs. 24 for CoNS). Hands-on time preparing DNA, carrying out the PCR, and evaluating results was less than 5 h.

CONCLUSIONS

The assay is largely automated, easy to adapt, and has been shown to be rapid and reliable. Fast detection and differentiation of S. aureus, CoNS, and the mecA gene by means of this real-time PCR protocol may help expedite therapeutic decision-making and enable earlier adequate antibiotic treatment.

摘要

背景

金黄色葡萄球菌长期以来一直被认为是一种主要病原体。耐甲氧西林金黄色葡萄球菌(MRSA)和耐甲氧西林表皮葡萄球菌(MRSE)是全球最常见的多重耐药病原体,经常引起医院感染和社区获得性感染。

方法

在本初步研究中,我们测试了一种聚合酶链反应(PCR)方法,以在临床环境中快速区分葡萄球菌并鉴定mecA基因。

结果

与传统微生物检测相比,实时PCR检测对金黄色葡萄球菌和凝固酶阴性葡萄球菌(CoNS)的检测率更高(金黄色葡萄球菌分别为55%对32%,CoNS为63%对24%)。制备DNA、进行PCR和评估结果的实际操作时间不到5小时。

结论

该检测方法在很大程度上实现了自动化,易于应用,并且已被证明快速可靠。通过这种实时PCR方案快速检测和区分金黄色葡萄球菌、CoNS和mecA基因,可能有助于加快治疗决策,并实现更早的充分抗生素治疗。

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本文引用的文献

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Rapid detection of methicillin-resistant Staphylococcus aureus from screening enrichment broths by real-time PCR.
Eur J Clin Microbiol Infect Dis. 2005 Jun;24(6):423-7. doi: 10.1007/s10096-005-1336-4.
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Methicillin-resistant staphylococci.
J Clin Pathol. 1961 Jul;14(4):385-93. doi: 10.1136/jcp.14.4.385.

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