聚合酶链反应反向膜杂交技术检测临床标本中结核分支杆菌的临床意义

Detection and clinical significance of mycobacterium tuberculosis by PCR-reverse cross-blot hybrid- ization method

目的探讨聚合酶链反应（ＰＣＲ）反向膜杂交技术捡测临床标本中结核分支杆酋（ＴＢ）ＤＮＡ的价值。方法用ＰＣＲ反向膜杂交技术检测 ５２２例结核及６０例非结核患者临床标本中的ＴＢ－ＤＮＡ，同时用培养、抗酸染色涂片及常规ＰＣＲ法作对照。结果ＰＣＲ反向膜杂交法阳性率为８０．６％（４１１／６９０），培养为１８．１（１２５／６９０），镜检为１３．９％（９６／６９０），常规ＰＣＲ为５９．６％（４１１／６９０），ＰＣＲ反向膜杂交法与常规法比较（Ｐ＜０．０００１），差异有显著意义。ＰＣＲ反向膜杂交法阳性检出率高于常规ＰＣＲ（Ｐ＞０．０５），但差异无显著意义；该技术假阳性为零。结论ＰＣＲ反向膜杂交技术检测临床标本中ＴＢ－ＤＮＡ具有快速、高度特异性和敏感性，可为结核病的临床早期诊断提供一种新的病原学诊断手段。

Objective To explore the value of detection of M. Tuberculosis in clinical specimens by PCR- reverse cross-blot hybridization method. Methods M. Tuberculosis DNA of clinical specimens in 522 patients with tuberculosis and 60 cases of nontuberculosis had detected by PCR-reverse cross-blot hybridization method, and compared with results smear fast-acid staining.culture and routine PCR from same 582 cases. Results In 522 pa- tients with tuberculesis, the positive rate of PCR-reverse cross-blot hybridization method was 80.6% routine PCR 59.6%,smear fast-acid staining 13.9% and culture 18.l%,respectively. The positive rates of PCR-re- verse cross-blot hybridization method were significantly higher than the smear and culture(P <0.001), the posi- tive rate of PCR-reverse cross-blot hybridization method was higher than that routine PCR(P > 0.05) . False positive rate of PCR-reverse cross-blot hybridization method was 0%. ConclusIon PCR-reverse cross-blot hybridization method is a rapid, simple, specific and sensitive method for detection of M. tuberculosis in clinical specimens and it has great value in the diagnosis and differentiation of tuberculous.