摘要目的 探讨应用荧光定量聚合酶链反应(qPCR)对疑似结核组织中结核分枝杆菌检测的可行性.方法 利用荧光qPCR对四川大学华西医院2011至2012年间1000例连续疑似结核组织中特异性结核分枝杆菌复合物IS6110 DNA片段进行扩增.齐-尼抗酸染色作为对照.扩增片段测序后与比对结核分枝杆菌基因序列比对验证其一致性.结果 1000例疑似结核组织中抗酸染色阳性513例,检出率为51.3％；荧光qPCR阳性546例,检出率54.6％.抗酸染色与荧光qPCR的符合率为73.1％.抗酸染色与荧光qPCR同时检测,可提高检出率14.4％.综合抗酸染色和qPCR分析结果,检出结核分枝杆菌外的抗酸染色阳性菌感染病例7例,其中包括3例麻风分枝杆菌感染,4例非结核分枝杆菌感染.结论 荧光qPCR对石蜡组织结核分枝杆菌检测敏感性略高于齐-尼抗酸特殊染色.同时应用两项检查可以提高检出率,并且可排除非结核分枝杆菌感染的漏诊.

abstractsObjective To investigate the feasibility of real-time fluorescent quantitative (qPCR)assay in detecting mycobacterium tuberculosis complex (MTB) in paraffin embedded tissues for diagnostic purpose.Methods Using qPCR assay,1000 consecutive formalin-fixed and paraffin embedded (FFPE) tissues (from 2011 to 2012) suspected of MTB infection were tested by amplifying the MTB specific insertion sequence 6110 (IS6110).The specificity of the PCR product was confirmed by Sanger sequencing as compared with the MTB genomic DNA of the IS6110 sequence.Tissues with Ziehl-Neelsen acid-fast staining were used as control.Results In the 1000 samples,513 were positive for mycobacterium by Ziehl-Neelsen acid-fast staining (detection rate 51.3％); whereas 546 were MTB positive by qPCR assay (detection rate 54.6％).Concordance rate for both assays was 73.1％.The diagnosis rate increased by 14.4％ by combinination of Ziehl-Neelsen acid-fast staining and qPCR results.More interestingly,by analyzing the Ziehl-Neelsen acid-fast staining and qPCR results three cases of M.leprae infection and four cases of nou-tuberculous Mycobacterium (NTM) infection were identified.Conclusions qPCR detection of MTB in FFPE tissue is more sensitive than Ziehl-Neelsen acid-fast staining assay.Combination of these two assays can increase the detection rate and also identify some rare cases of NTM infection.