摘要目的 评估RNA恒温扩增实时荧光检测技术(SAT)在临床痰液标本结核分枝杆菌检测中的价值.方法 对2011年9至12月上海市肺科医院230例临床确诊肺结核患者和78例其他呼吸系统疾病患者的痰液分别用SAT法、BD960培养法、罗氏培养法和集菌涂片法同时进行检测.SAT法与BD960培养法结果不符标本,再用结核分枝杆菌核酸扩增荧光体外检测试剂盒(PCR-TB)进行检测,同时对BD960培养阳性菌株和SAT扩增产物进行扩增及测序,对菌种进行鉴定.采用x2检验比较SAT法与上述3种方法对结核病患者的阳性检出率.结果 以BD960培养法作为金标准(剔除BD960培养法中有7份污染菌),则SAT方法的敏感度为90.5％( 95/105),特异度为84.2％( 165/196),阳性预测值为75.4％ (95/126),阴性预测值为94.3％( 165/175).SAT法与BD960培养法结果符合率为86.4％( 260/301).223份临床确诊的肺结核患者痰标本SAT法、BD960培养法、罗氏培养法和涂片法的阳性检出率分别为56.5％( 126/223)、45.7％( 102/223)、41.7％( 93/223)和37.2％( 83/223),SAT法显著高于其他3种方法,差异有统计学意义(x2=4.087,P＜0.05).结论 SAT技术用于结核分枝杆菌实验室诊断具有特异度高、敏感度好,操作简便、快速,污染率低的特点,有望成为实验室诊断的新方法.

abstractsObjective To evaluate the clinical value of the isothermal RNA amplification assay (SAT) for detection of Mycobacterium tuberculosis in sputum samples.Methods Sputum specimens from 230 patients with diagnosed tuberculosis and 78 cases of other respiratory diseases during September to December 2011 were detected using SAT,BD960 culture,LowenStein-Jensen( L-J ) culture and concentrated smear simultaneously.The samples with different results between SAT and BD960 culture were tested by Mycobacterium tuberculosis PCR fluorescence diagnosis kits.Strains were identified by amplification and sequencing the BD960 culture-positive isolates and SAT amplification products.Positive detection rate of SAT and other three methods for patients with tuberculosis were compared by chi-square test.Results Using the results of BD960 culture as the golden standard (7 cases of pollution bacteria in BD960 culture was rejected ),the sensitivity,specificity,positive predictive value,and negative predictive value of SAT was 90.5％ (95/105),84.2％ (165/196),75.4％ (95/126),94.3％ (165/175),respectively.The agreement rate of SAT and BD960 culture was 86.4％ (260/301).For 223 tuberculosis patients,the positive detection rate of SAT,BD960 culture,L-J culture and concentrated smear was 56.5％ ( 126/223 ),45.7％ ( 102/223 ),41.7％ ( 93/223 ) and 37.2％ ( 83/223 ) respectively.The positive detection rate of SAT is significantly higher than the other three methods (x2 =4.087,P ＜ 0.05 ).Conclusion SAT,as a new technology for laboratory diagnosis of TB,has high specificity and sensitivity.The operation is fast and simple,and the pollution rate is low.It is a promising laboratory diagnosis method.