聚合酶链反应-增强化学发光检测结核分枝杆菌

Detection of Mycobacterium tuberculosis by PCR-enhanced chemiluminescence assay

目的 探讨微孔板杂交技术结合增强化学发光 (ECL)检测结核分枝杆菌 (MTB)。方法 同时应用聚合酶链反应 (PCR) ECL、PCR 比色法与PCR电泳检测 6 0例活动性肺结核和 5 4例健康对照痰标本 ,比较结果。结果 PCR ECL、PCR 比色法与PCR电泳总阳性率分别为 76 .7%、6 3.3%、5 0 .0 % ,其中PCR ECL阳性率显著高于PCR电泳 (P <0 .0 5 ) ;对涂片阳性标本 ,阳性率分别为 85 .7%、85 .7%、71.4% ,差异无显著性 (P >0 .0 5 ) ;对涂片阴性标本 ,阳性率分别为 73.9%、5 6 .5 %、43.5 % ,PCR ECL阳性率高于PCR电泳 (P <0 .0 5 )。另外PCR ECL、PCR 比色法与PCR电泳特异性分别为 92 .6 %、96 .3%、96 .3% ,差异无显著性 (P >0 .0 5 )。结论 PCR ECL灵敏度高 ,能提高临床标本MTB检出率 ,尤其是对涂片阴性痰标本 ,并且该方法操作简单 ,结果客观。

Objective To evaluate the value of microplate hybridization assay and PCR enhanced chemiluminescence (ECL) in the detection of Mycobacterium tuberculosis . Methods 60 sputum specimens from patients with active pulmonary tuberculosis and 54 sputum specimens from people without tuberculosis were tested by PCR ECL, PCR colorimetry and PCR agarose gel electrophoresis respectively. The results were compared with each other. Results The positive rates of PCR ECL, PCR colorimetry and PCR agarose gel electrophoresis were 76.7%,63.3% and 50.0% respectively. There was a significant difference between the positive rates of PCR ECL and PCR agarose gel electrophoresis( P < 0.05 ). In the smear positive specimens, there was no significant difference among the positive rates of three methods( P >0.05). In the smear negative specimens, the positive rates of three methods were 73.9%,56.5% and 43.5% respectively. There was a significant difference between the positive rates of PCR ECL and PCR agarose gel electrophoresis( P <0.05). The specificity of PCR ECL, PCR colorimetry and PCR agarose gel electrophoresis were 92.6%, 96.3 % and 96.3% respectively. There was no significant difference among them( P > 0.05 ). Conclusion PCR ECL was sensitive and useful, especially in the detection of smear negative specimens. Furthermore, the method was easily performed and the interpretation of results was objective.