中华结核和呼吸杂志
中華結覈和呼吸雜誌
중화결핵화호흡잡지
Chinese Journal of Tuberculosis and Respiratory Diseases
2013年
3期
198-201
,共4页
分枝杆菌,结核%抗药性%吡嗪酰胺%DNA突变分析
分枝桿菌,結覈%抗藥性%吡嗪酰胺%DNA突變分析
분지간균,결핵%항약성%필진선알%DNA돌변분석
Mycobacterium tuberculosis%Drug resistance%Pyrazinamide%DNA mutational analysis
目的 应用高分辨率熔解曲线(HRM)技术建立快速检测MTB对吡嗪酰胺的耐药性.方法 收集深圳市慢性病防治中心2007-2009年耐药监测项目中的临床分离菌株,用基因测序法检测pncA基因突变情况,选取49株突变型菌株和78株野生型菌株作为研究对象,建立HRM检测pncA基因突变方法,并应用于吡嗪酰胺耐药性评估.以BACTEC MGIT 960结果为金标准,评价HRM的应用价值.结果 49株突变型菌株中HRM检测出48株突变,1株双突变位点误判为野生型,78株野生型菌株中HRM检测出6株突变型.BACTEC MGIT 960药敏方法测定结果,127株临床分离株中吡嗪酰胺耐药62株,敏感65株；HRM检测62株耐药株中有53株耐药,65株敏感株中有64株敏感.以BACTEC MGIT 960为药物敏感性的金标准,其敏感度为85.5％ (53/62),特异度为98.5％(64/65),Youden指数为0.840.结论 HRM检测吡嗪酰胺耐药性与BACTEC MGIT 960法有很好的一致性,具有快速、简便的优点,有较好的临床应用前景.
目的 應用高分辨率鎔解麯線(HRM)技術建立快速檢測MTB對吡嗪酰胺的耐藥性.方法 收集深圳市慢性病防治中心2007-2009年耐藥鑑測項目中的臨床分離菌株,用基因測序法檢測pncA基因突變情況,選取49株突變型菌株和78株野生型菌株作為研究對象,建立HRM檢測pncA基因突變方法,併應用于吡嗪酰胺耐藥性評估.以BACTEC MGIT 960結果為金標準,評價HRM的應用價值.結果 49株突變型菌株中HRM檢測齣48株突變,1株雙突變位點誤判為野生型,78株野生型菌株中HRM檢測齣6株突變型.BACTEC MGIT 960藥敏方法測定結果,127株臨床分離株中吡嗪酰胺耐藥62株,敏感65株；HRM檢測62株耐藥株中有53株耐藥,65株敏感株中有64株敏感.以BACTEC MGIT 960為藥物敏感性的金標準,其敏感度為85.5％ (53/62),特異度為98.5％(64/65),Youden指數為0.840.結論 HRM檢測吡嗪酰胺耐藥性與BACTEC MGIT 960法有很好的一緻性,具有快速、簡便的優點,有較好的臨床應用前景.
목적 응용고분변솔용해곡선(HRM)기술건립쾌속검측MTB대필진선알적내약성.방법 수집심수시만성병방치중심2007-2009년내약감측항목중적림상분리균주,용기인측서법검측pncA기인돌변정황,선취49주돌변형균주화78주야생형균주작위연구대상,건립HRM검측pncA기인돌변방법,병응용우필진선알내약성평고.이BACTEC MGIT 960결과위금표준,평개HRM적응용개치.결과 49주돌변형균주중HRM검측출48주돌변,1주쌍돌변위점오판위야생형,78주야생형균주중HRM검측출6주돌변형.BACTEC MGIT 960약민방법측정결과,127주림상분리주중필진선알내약62주,민감65주；HRM검측62주내약주중유53주내약,65주민감주중유64주민감.이BACTEC MGIT 960위약물민감성적금표준,기민감도위85.5％ (53/62),특이도위98.5％(64/65),Youden지수위0.840.결론 HRM검측필진선알내약성여BACTEC MGIT 960법유흔호적일치성,구유쾌속、간편적우점,유교호적림상응용전경.
Objective To evaluate the application of a high-resolution melting (HRM) curve analysis for rapid detection of pyrazinamide resistance in Mycobacterium tuberculosis (M.tuberculosis).Methods Clinical isolates of M.tuberculosis were collected from Shenzhen from 2007-2009 and pncA mutations were characterized by sequence analysis.HRM analysis method was established and used to determine pncA gene mutation in 49 pncA mutant isolates and 78 wild type isolates.BCATEC MGIT 960 was used as the standard method to evaluate application of the HRM curve.Results Among 49 pncA mutant isolates,48 isolates were characterized as mutant and 1 isolate with double point mutations misclassified as wild type by using HRM detection.In 78 wild type isolates,HRM found 6 mutant isolates.Using the BCATEC MGIT 960 to detect pyrazinamide drug susceptibility,62 isolates were found resistant and 65 were susceptible.Among the 62 resistant strains by HRM results 53 strains were resistant,and among the 65 susceptible stains by HRM results 64 were susceptible.Compared to the results of BACTEC MGIT 960,the sensitivity and specificity of HRM were 85.5％ (53/62) and 98.5％ (64/65),respectively.The Youden index was found to be 0.840.Conclusions The results showed a good correlation between HRM curve analysis and BACTEC MGIT 960.HRM curve analysis is a rapid and accurate method that can be used as a screening test for identification of resistance to pyrazinamide.
