【摘要】 目的建立多重实时荧光PCR方法对医源性感染标本中的耐甲氧西林葡萄球菌进行快速检测,为院内葡萄球菌感染的监测提供技术方案。方法通过对金黄色葡萄球菌的耐热核酸酶基因片段(nuc)、葡萄球菌共有的基因片段(tuf)和甲氧西林耐药性基因片段(mecA)的序列进行比对,设计能同时检测金黄色葡萄球菌和葡萄球菌属及其耐药基因的引物和探针序列,通过检测标准菌株和实际采集标本,对方法进行评估。结果 nuc和tuf片段设计采用标准菌株验证方法准确性,未发现假阳性和假阴性结果;mecA片段与分离培养鉴定检测结果一致;对于混合标本,应结合PCR法和分离培养法的结果,对病原菌耐药状况进行具体分析。结论该方法检测时间短,适用于医疗机构对医源性感染的快速分析。更多还原

【Abstract】 Objective To establish multiplex realtime fluorescence polymerase chain reaction(PCR)for the detection of methicillin-resistant Staphylococcusin nosocomial infection samples to provide fast and accurate method to monitor nosocomial infection.Methods Alignment of tufand mecAof Staphylococcus and nuc of Staphylococcus aureus was performed to design primer and probe labeled with FAM/HEX/ROX to detect strains and nosocomial samples.Differences of culture and PCR detection results were analyzed.Results Nuc and tufrealtime PCR was verified by detecting strains.MecArealtime PCR was agreement with culture method.It was necessary to anatomize details of realtime PCR and culture results for the detection of mixed samples.Conclusion Multiplex realtime PCR could be applied for rapid analysis of nosocomial infection.更多还原