聚合酶链反应快速诊断结核病的应用研究  

STUDY ON RAPID DIAGNOSIS OF TUBERCULOSIS BY POLYMERASE CHAIN REACTION

作　　者：唐小平[1] 罗惠娟[1] 黄已实[1] 卢群馨 

机构地区：[1]广州市传染病医院

出　　处：《中华微生物学和免疫学杂志》1994年第1期26-28,共3页Chinese Journal of Microbiology and Immunology

基　　金：广州市医药卫生青年基金

摘　　要：应用聚合酶链反应（ＰＣＲ）技术建立了扩增结核杆菌特异重复序列ＩＳ６１１０部分基因的方法。对９种抗酸分枝杆菌、３种普通菌进行检测，结果仅人型结核杆菌、牛型结核杆菌及ＢＣＧ扩增出１２３ｂｐ特异性条带，ＰＣＲ产物经ＳａｌⅠ酶切后产生７０ｂｐ与５３ｂｐ两个片段。ＰＣＲ检测人型结核杆菌的敏感性达１０ｆｇＤＮＡ或５个菌体。应用ＰＣＲ检测了１０９份结核临床标本，其总阳性率为７２．５％，明显高于抗酸染色涂片（２．８％）与细菌培养（９．２％）的阳性率（Ｐ＜０．００１）。ＰＣＲ的检出率也较ＥＬＩＳＡ法检测抗ＰＰＤ－ＩｇＧ的阳性率（６３．３％）为高，但尚无统计学差异（ｐ＞０．０５），但是ＥＬＩＳＡ检测抗体有１９．０％的假阳性。研究表明，ＰＣＲ是一种特异、敏感、快速诊断结核病的方法。An assay of amplification for a 123bp segment derived from the repetitive DNA IS6110 specific for Mycobacte-ria tuberculosis complex was developed by using polymerase chian reaction(PCR).9 strains of acid-fast mycobacte-ria and 3 strains of non-mycobacteria were tested, and the specific PCR product was obtained only from M. tubercu-losis,M. bovis and M. bovis BCG. Through digestion of endonuclease Sal Ⅰthe product was separated into twofragments,70bp and 53bp. The assay could detect as less as 10fg DNA of M·tuberculosis(or 5 organisms).It was used to detect M.tuber-culosis DNA in 109 clinical specimens from patients with tuberculosis(66 pleural fluid aspirates and 43 cere-brospinal fluids ),and it was shown that the positivity rate of PCR(72.5%)was significantly higher than that ofconventional acid-fast stain(2.8%)and culture techniques(9.2%).The positivity rate of PCR was also higherthan that of ELISA for detection of Anti-PPD IgG antibody(63.3%),but the difference didn't reach statisticalsignificance. But there were 19.0%false-positives in anti-PPD IgG antibody detection. Our results suggest thatPCR may be a specific, sensitive and rapid technique for the diagnosis of tuberculosis。

关 键 词：聚合酶链反应 结核病 分枝杆菌 

分 类 号：R520.4[医药卫生—内科学]