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目的 应用敏感特异的PCR反向膜探针杂交ELISA法 (以下称PCR杂交梳 )检测结核杆菌 ,并对该法与传统的结核菌检测方法进行比较评价。方法 用PCR杂交梳和涂片抗酸染色 ,结核菌培养 ,PCR凝胶电泳法分别检测 140例临床标本 ,进行比较分析 ,并用相关的微生物对PCR杂交梳技术进行特异性和敏感性试验。结果 97例已确诊结核病的标本 ,经涂片抗酸染色 ,结核菌培养 ,PCR凝胶电泳和PCR杂交梳法检测结核杆菌 ,其阳性率分别为 6 .2 %、14.4%、5 7.7%、85 .6 % ,43例非结核肺病标本经上述几种方法检测结核菌 ,阳性率除PCR凝胶电泳法为为 13.9% ,其它方法均未检出 ,TB -DNA阳性的PCR产物经 1∶10 4稀释 ,用反向膜探针杂交ELISA法检测仍有可见反应并与相关微生物无交叉反应。结论 PCR杂交梳检测结核杆菌 ,快速准确 ,灵敏度高 ,特异性强 ,克服了以往结核菌常规检验方法 ,费时过长 ,检出率低及PCR凝胶电泳法易引起假阳性污染等缺点 ,结果真实可靠 ,对结核病的临床诊断 ,治疗方案选择和疗效观察有较大的指导意义
Objective To detect Mycobacterium tuberculosis by ELISA-specific reverse-membrane hybridization ELISA (hereinafter referred to as PCR hybridization comb), and compare the method with the traditional detection methods of Mycobacterium tuberculosis. Methods 140 cases of clinical samples were detected by PCR hybridization comb, acid-fast staining with smear, tubercle bacilli culture and PCR gel electrophoresis respectively. The clinical and pathological characteristics of 140 clinical samples were compared and analyzed by PCR-specific combing technique. Results A total of 97 cases of confirmed tuberculosis were detected by smear antacid staining, Mycobacterium tuberculosis culture, PCR gel electrophoresis and PCR hybridization comb. The positive rate of Mycobacterium tuberculosis was 6.2%, 14.4% and 5 7.7%, respectively. 85. 6%, 43 cases of non-tuberculosis lung specimens by the above methods to detect TB, the positive rate in addition to PCR gel electrophoresis was 13.9%, and other methods were not detected, TB-DNA-positive PCR products by 1: 10 4 diluted with reverse membrane probe hybridization ELISA still visible reaction and no cross-reaction with the relevant microorganisms. Conclusion PCR hybridization comb detection Mycobacterium tuberculosis, rapid and accurate, high sensitivity and specificity, to overcome the previous routine testing methods of TB, over time, the detection rate is low and PCR gel electrophoresis easily lead to false positives and other shortcomings, the results True and reliable, the clinical diagnosis of tuberculosis, treatment options and curative effect observation has a greater guiding significance