直接杂交法检测痰液中结核分枝杆菌  

Detection of clinical sputum specimens of Mycobacterium tuberculosis by direct hybridization

作　　者：杨华卫[1] 杨树德[1] 庄玉辉[2] 张灵霞[2] 

机构地区：[1]卫生部临床检验中心,北京100730 [2]中国人民解放军第三零九医院,北京100091

出　　处：《临床检验杂志》2001年第1期1-3,共3页Chinese Journal of Clinical Laboratory Science

基　　金：国家自然科学基金资助项目!(编号:39670696)

摘　　要：目的建立了一种高度灵敏、特异、快速和简便的探针杂交检测技术，以解决临床上结核病快速诊断的难题。方法采用简易的方法处理标本，以对结核分枝杆菌特异的生物素标记 １８８ ｈｐ ＤＮＡ探针进行点杂交，与链霉亲合素标记的辣根过氧化酶偶联后，以化学发光自显影方式记录检测信号。结果 此法的敏感性可检出２００个菌，较ＰＣＲ法为高。对２０种分枝杆菌和９种非分枝杆菌检测时发现只有结核分枝杆菌呈阳性。以本法检测１０４例临床标本，结果发现，对其中４５例结核病人痰涂片阳性标本，４１例结核病人痰涂片阴性标本和１８例非结核标本，检测阳性率分别为１００％、６５．９％和０．０％。结论本法简便，特异。Objective A new highly sensitive, specific, fast and simple hybridization method was established for rapid diagnosis of tubercu- losis. Methods Samples were processded by a simple method, dot blotted and hybridized with a biotin-labeled 188 bp DNA probe specific to Mycobaterium tuberculosis, and detected by auto-chehiluminograghy.Results For direct hybridization method, 200 organisms were able to be detected, offering a higher sensitivity than PCR. The specificity was excellent: only M. tuberculosis were positive among 20 mycobacterium and 9 no-mycobacterium. Total 104 clinical samples were tested, including 45 sputum smear positive, 4 1 negative specimens in pulmonary tuberculosis and 18 non - mycobacterium ones. The positive rates of then were l00%, 65.9% and 0.0% by 188 bp probe and auto-chemiluminography respectively. Conclusion This mothod is a simple and specificity.

关 键 词：结核分枝杆菌 化学发光自显影 DNA探针点杂交 痰液 

分 类 号：R446.5[医药卫生—诊断学] R378.911[医药卫生—临床医学]