摘要
目的探讨荧光聚合酶链反应(PCR)熔解曲线法快速检测结核分枝杆菌复合群(MTBC)及异烟肼耐药突变的临床价值。方法对延安市传染病医院2018年4-12月住院的370例肺结核患者痰标本采用MGIT320进行MTBC培养及药敏试验,同时应用荧光PCR熔解曲线法进行MTBC和异烟肼耐药突变检测,对检出MTBC的差异进行分析,以培养法药敏为金标准,评估熔解曲线法耐药突变检测的灵敏度、特异度、一致性等。结果370例患者培养法检出MTBC175株,阳性率47.3%,熔解曲线法检出157株,阳性率42.4%,两种方法检测的结果差异无统计学意义(P=0.073),一致性中等(Kappa=0.510);同时与有异烟肼药敏结果的115株进行比较,熔解曲线法灵敏度95.0%,特异度93.7%,与培养法结果比较差异无统计学意义(P=0.125),具有高度一致性(Kappa=0.807)。结论熔解曲线法试剂盒可用于结核菌快速筛查及耐药突变检测,对异烟肼耐药突变检测具有高灵敏度和特异度,有助于缩短耐药结核病诊断时间。
Objective To evaluate the clinical utility of fluorescence PCR melting curve analysis in rapid detection of Mycobacterium tuberculosis complex (MTBC) and isoniazid resistant gene mutations.Methods The 370 patients with pulmonary tuberculosis treated in Yan'an Infectious Diseases Hospital from April to December 2018 were included in this study.The sputum specimens were tested with MGIT320 to isolate MTBC and conduct antimicrobial susceptibility testing.Meanwhile,fluorescence PCR melting curve analysis was also used to detect MTBC and isoniazid resistance mutations.The results achieved by the two methods were compared in terms of MTBC detection,as well as the sensitivity,specificity and consistency in identification of resistant gene mutations,taking culture and antimicrobial susceptibility testing as gold standard.Results Overall,175 (47.3%) strains of MTBC were detected by culture method,and 157 (42.4%) strains were detected by melting curve method (P=0.073).The results were consistent (kappa=0.510).A total of 115 strains had isoniazid susceptibility data.The sensitivity and specificity of the melting curve method in identifying isoniazid susceptibility were 95.0% and 93.7%,respectively when compared with the results of culture method (P=0.125).Both methods showed high consistency (kappa=0.807).Conclusions The melting curve analysis kit can be used for rapid screening of tuberculosis and detection of isoniazid resistant gene mutations,showing high sensitivity and specificity.It is helpful to reduce the time to diagnosis of drugresistant tuberculosis.
作者
阳央
冯智
侯欢
李芳芹
YANG Yang;FENG Zhi;HOU Huan;LI Fangqin(Yan'an University Medical College,Yan'an Shaanxi 716000,China)
出处
《中国感染与化疗杂志》
CAS
CSCD
北大核心
2019年第4期405-409,共5页
Chinese Journal of Infection and Chemotherapy
关键词
结核分枝杆菌
熔解曲线法
快速检测
异烟肼耐药突变
Mycobacterium tuberculosis
melting curve analysis
rapid detection
isoniazid resistance mutation
