CAJ | 학술논문

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。
목적：건립다중실시형광PCR방법대의원성감염표본중적내갑양서림포도구균진행쾌속검측，위원내포도구균감염적감측제공기술방안。방법통과대금황색포도구균적내열핵산매기인편단（nuc）、포도구균공유적기인편단（tuf）화갑양서림내약성기인편단（mecA）적서렬진행비대，설계능동시검측금황색포도구균화포도구균속급기내약기인적인물화탐침서렬，통과검측표준균주화실제채집표본，대방법진행평고。결과 nuc화tuf 편단설계채용표준균주험증방법준학성，미발현가양성화가음성결과；mecA편단여분리배양감정검측결과일치；대우혼합표본，응결합PCR법화분리배양법적결과，대병원균내약상황진행구체분석。결론해방법검측시간단，괄용우의료궤구대의원성감염적쾌속분석。
Objective To establish multiplex realtime fluorescence polymerase chain reaction (PCR)for the detection of methi-cillin-resistant Staphylococcus in nosocomial infection samples to provide fast and accurate method to monitor nosocomial infection. Methods Alignment of tuf and mecA of 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 tuf realtime PCR was verified by detecting strains.MecA realtime PCR was agreement with cul-ture method.It was necessary to anatomize details of realtime PCR and culture results for the detection of mixed samples.Conclu-sion Multiplex realtime PCR could be applied for rapid analysis of nosocomial infection.