PCR与单克隆抗体ELESA夹心法快速诊断结核病的研究

应用聚合酶性反应（ＰＣＲ）技术建立的扩增结核杆菌复合体特异重复序列ＩＳ９８６基因的方法和用单克隆抗体ＴＢ１５－Ｃ３经ＥＬＩＳＡ夹心法检测结核杆菌特异性抗原决定簇，对１０种抗酸杆菌，２种普通菌进行了检测．ＰＣＲ仅对结核杆菌复合体扩增出２４５ｈｐ特异性条带，ＥＬＩＳＡ检测除人型结核杆菌、ＢＣＧ阳性外，还与鸟型及瘰疬分枝杆菌反应阳性．ＰＣＲ检测人型结核杆菌的敏感性为１ｐｇ，相当于１３个左右细菌，ＥＬＩＳＡ检测的被感性为１５ｎｇ／ｍｌ．应用ＰＣＲ及ＥＬＩＳＡ检测了９６份结核临床标本．ＰＣＲ的检出率高于抗酸染色涂片的阳性率（Ｐ＜０．０５），ＰＣＲ的检出率虽高于细菌培养的阳性率，但相差不显著（Ｐ＞０．０５）．ＥＬＩＳＡ检测阳性率明显高于抗酸染色涂片、细菌培养和ＰＣＲ的阳性率（Ｐ＜０．００１）．ＥＬＩＳＡ的假阳性率高于ＰＣＲ的假阳性率，但相差不显著（Ｐ＞０．０５）．研究表明，ＰＣＲ及ＥＬＩＳＡ均是特异、敏感、快速的诊断结核病的方法．但在使用中又各自有其特点．

Comparison of PCR and ELISA on Rapid Diagnois of Tuberculosis

An assay of amplincation for a 245hp segment derived from the repetitive DNA IS986 specific for Mycobacterium tuberculosis complex was developed by using Polymerase chain reaction(PCR),and a sandwich enzyme linked immunosorbant assay (ELISA) was developed by usingmonoclonal antibody TB15-C3 that recognized specific antigenic determinant of Mycobacterium tuberculosis. Ten strains of acid-fast mycobacteria and 2 strains of non-mycobacteria were tested.Thespecific PCR product was obtained only from Mycobacterium tuberculosis complex. Besides M. tuberculosis and BCG, M. avium and M.scrofulaceum was POsitive by ELISA analysis. The sensitivity ofdetection of M.tuberculosis genomic DNA and bacteria suspension by PCR was lpg or 13 viable bacteria cells respeCtively.The sensitivity of detection of PPD by ELISA was 15 ng / ml. They were usedto detect M.tuberculosis DNA and specific antigenic determinant in 96 clinical specimens from patients with tuberculosis. It was shown that the positivity rate of PCR was higher than that ofconventional acid-fast stain(P<0.05) and was not remarkably higher than that of culturgtechniques (P< 0.05).The POsitivity rate of ELISA for detection of speCific antigenic detendnant w4smuch higher than that of conventional acid--fast stain, culture techniques and PCR(P<0.01).The falsePOsitly rate of ELISA was not remerkably higher than that of PCR (P<0.05).Our resultS suggest thatPCR and ELISA may be speCieic, sensitive and rapid techniques for ti.,e diagnosis of tuberculosis.