CAJ | 학술논문

目的评价探针熔解分析法快速检测结核分枝杆菌乙胺丁醇耐药突变的应用价值,为临床检测提供指导依据.方法 613份痰标本于2009年9月至2010年4月收集自厦门市疾病预防控制中心、厦门市第一医院和漳州市疾病预防控制中心.结核分枝杆菌H37Rv标准株来源于国家结核病参比实验室.37株包含结核与非结核分枝杆菌的标准盘由中国药品生物制品检定所提供.首先采用梯度稀释的野生型标准株H37Rv DNA考察探针熔解曲线分析法的分析灵敏度,然后用标准盘验证其检测特异性,最后以测序法为对照方法,采用613份结核分枝杆菌的标本评价该方法的临床应用价值.结果探针熔解分析法可检测到3拷贝/反应,且能特异检测结核分枝杆菌.613份结核分枝杆菌的检测结果显示,符合要求的583份标本的试剂盒检测结果与测序结果完全一致,其中embB306突变菌株数为34株,embB 378-380突变菌株数为23株,embB 406突变菌株数为3株,embB 497突变菌株数为3株.结论探针熔解分析法检测结核分枝杆菌乙胺丁醇耐药突变特异性好,灵敏度高,能有效地检测临床标本常见乙胺丁醇耐药突变位点,是值得推广的快速检测方法.
목적 평개탐침용해분석법쾌속검측결핵분지간균을알정순내약돌변적응용개치,위림상검측제공지도의거.방법 613빈담표본우2009년9월지2010년4월수집자하문시질병예방공제중심、하문시제일의원화장주시질병예방공제중심.결핵분지간균H37Rv표준주래원우국가결핵병삼비실험실.37주포함결핵여비결핵분지간균적표준반유중국약품생물제품검정소제공.수선채용제도희석적야생형표준주H37Rv DNA고찰탐침용해곡선분석법적분석령민도,연후용표준반험증기검측특이성,최후이측서법위대조방법,채용613빈결핵분지간균적표본평개해방법적림상응용개치.결과 탐침용해분석법가검측도3고패/반응,차능특이검측결핵분지간균.613빈결핵분지간균적검측결과현시,부합요구적583빈표본적시제합검측결과여측서결과완전일치,기중embB306돌변균주수위34주,embB 378-380돌변균주수위23주,embB 406돌변균주수위3주,embB 497돌변균주수위3주.결론 탐침용해분석법검측결핵분지간균을알정순내약돌변특이성호,령민도고,능유효지검측림상표본상견을알정순내약돌변위점,시치득추엄적쾌속검측방법.
Objective To evaluate the potential use of a probe melting analysis (PMA) assay in detecting the embB mutations which confer resistance against ethambutol in Mycobacterium tuberculosis. Methods The analysis sensitivity and specificity of PMA were investigated by detecting a serially diluted H37 Rv DNA and a reference panel from National Institute for the Control of Pharmaceutical and Biological Product. Six hundred and thirteen sputum samples were collected from the Xiamen Center for Disease Control and Prevention, Xiamen First Hospital and Center for Zhangzhou Disease Control and Prevention from September 2009 to April 2010. The PMA assay was then evaluated by detecting 613 clinical isolates and the results were compared with the sequencing results. Results The PMA assay could specifically detect Mycobacterium tuberculosis and had a limit of detection of 3 copies per reaction. The assay results with 613 clinical isolates showed that PMA gave a 100% concordance with sequencing in the 583 qualified samples, among which 34 were mutations at embB 306,23 at embB 378-380, 3 at embB 406 and 3 at embB 497. Conclusions PMA assay is a sensitive and specific method enabling efficient detection of common embB mutations causing ethambutol-resistance. The rapidness of this method together with its reliability would facilitate its use in routine testing.