聚合酶链反应-微孔板杂交法检测结核杆菌的临床应用及其评价

Clinical Application and Evaluation of Polymerase Chain Reaction Microwell Plat e Hybridization Assay in the Detection of Mycobacterium tuberculosis

目的 对聚合酶链反应 ( PCR) -微孔板杂交法检测结核杆菌的临床适用性和可行性进行评价。方法 收集 1130份临床标本 ,分别用抗酸染色、培养、PCR电泳及 PCR微孔板杂交法进行结核杆菌检测 ,并结合临床诊断和疗效观察对实验结果进行分析。结果 10 0份临床证实无结核病的体液标本经抗酸染色和 PCR电泳 ,分别有 1例和 2例假阳性 ,但经培养和 PCR微孔板杂交法未查出阳性 ;其余 10 30份高度怀疑含有结核杆菌的体液标本的检测结果 ,以 PCR微孔板杂交阳性检出例数最高 ( 481/ 10 30 ) ,其次为 PCR电泳 ( 40 6 / 10 30 )、培养 ( 36 5 /10 30 )以及抗酸染色 ( 2 5 6 / 10 30 ) ;χ2 检验结果显示 ,PCR微孔板杂交的结核杆菌阳性检出例数与其它三种方法比较均呈显著或极显著性差异 ( P<0 .0 0 83及 P<0 .0 0 17)。结论 PCR-微孔板杂交方法是特异、灵敏、准确、快速的结核杆菌检测方法 。

Objective To evaluate the clinical application an d feasibility of polymerase chain reaction microwell plate hybridization assay in the detection of M ycobacterium tuberculosis. Methods 1130 specimens with strong s uspicion for myc obacterium tuberculosis were collected from the hospitals and were detected by fast bacilli stain,culture,PCR electrophoresis and PCR microwell plate hybridi za tion respectively. The laboratory results were analyzed in combination with the symptoms and signs of patients and the observations on treatment.Also detected were 100 samples from the clinically evidenced non tuberculosis patients. Results In the 100 samples collected from the patients without tuber culosis, the PCR h ybridization method and culture method did not detect Mycobacterium tuberculosis , but the fast bacilli stain method and PCR electrophoresis method brought out o ne and two false positive results respectively. The results of testing the 1030 clinical samples which probably contained Mycobacterium tuberculosis demonstrate d that the PCR hybridization method had the highest positive rate (481/1030) am o ng the four methods, and the positive rates of the other three methods were PCR electrophoresis (406/1030), culture(365/1030) and fast bacilli stain(256/1030) i n proper order. The chi square test showed that there were significant differen c e between the PCR hybridization method and the other three methods respectively (P <0.0083 or P <0.0017).Conclusion PCR Hybridization m ethod is specific, sensitive,accurate and fast in detecting mycobacterium tuberc ulosis; it is worthy to be used clinically.