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首页> 外文期刊>Journal of Clinical Microbiology >Detection of Methicillin-Resistant Staphylococcus aureus and Simultaneous Confirmation by Automated Nucleic Acid Extraction and Real-Time PCR
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Detection of Methicillin-Resistant Staphylococcus aureus and Simultaneous Confirmation by Automated Nucleic Acid Extraction and Real-Time PCR

机译：耐甲氧西林金黄色葡萄球菌的检测以及自动核酸提取和实时PCR的同时确认

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摘要

A molecular assay for the simultaneous detection of a Staphylococcus aureus-specific gene and the mecA gene, responsible for the resistance to methicillin in staphylococci, was evaluated. The assay included an automated DNA extraction protocol conducted with a MagNA Pure instrument and real-time PCR conducted with a LightCycler instrument. The performance and robustness of the assay were evaluated for a suspension of methicillin-resistant S. aureus (MRSA) strain with a turbidity equivalent to a McFarland standard of 0.5, which was found to be the ideal working concentration. The specificity of the new molecular assay was tested with a panel of 30 gram-negative and gram-positive bacterial strains other than MRSA. No cross-reactivity was observed. In a clinical study, 109 isolates of MRSA were investigated. All clinical MRSA isolates gave positive results for the S. aureus-specific genomic target, and all but one were positive for the mecA gene. In conclusion, the new molecular assay was found to be quick, robust, and laborsaving, and it proved to be suitable for a routine molecular diagnostic laboratory.
机译：评价了一种同时检测金黄色葡萄球菌特异基因和 mecA 基因的分子检测方法，该基因负责葡萄球菌对甲氧西林的耐药性。该测定包括使用MagNA Pure仪器进行的自动DNA提取方案和使用LightCycler仪器进行的实时PCR。评估了耐甲氧西林 S悬浮液的测定性能和稳健性。金黄色葡萄球菌（MRSA）的浊度相当于McFarland标准的0.5，这是理想的工作浓度。用一组除MRSA以外的30克阴性和革兰氏阳性细菌菌株测试了新的分子测定的特异性。没有观察到交叉反应。在一项临床研究中，研究了109株MRSA分离株。所有临床MRSA分离株对 S均给出阳性结果。金黄色葡萄球菌特有的基因组靶标，除 mecA 基因外，其余所有均为阳性。总而言之，新的分子测定方法被发现具有快速，稳定和省力的特性，并且被证明适用于常规的分子诊断实验室。

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