结核病实验室3种快速检测方法的评价

Evaluation of three rapid methods in the detection of mycobacterium tuberculosis

  • 摘要: 用聚合酶链反应(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(McAb)TB15-C_3经酶联免疫吸附测定(ELISA)夹心法检测结核杆菌特异性抗原决定簇,以及用抗酸染色的方法,对10种抗酸杆菌,2种非分枝杆菌进行检测。PCR仅对结核杆菌复合体扩增出245bp特异性条带,McAb-EItISA检测除人型结核杆菌,BCG阳性外,还与鸟型及瘰疲分枝杆菌反应阳性,抗酸染色则所有分枝杆菌均呈阳性。PCR检测人型结核杆菌的敏感度为15ng/ml。抗酸染色敏感度为1×(104~105)个细菌。应用抗酸染色,PCR及McAb-ELISA检测了138份结核临床标本,PCR的检出率高于抗酸染色徐片的阳性率(P<0.05),McAb-ELISA检测阳性率明显高于抗酸染色涂片和PCR的阳性率(P<0.01)。研究表明,抗酸染色,PCR及ELISA均为快速结核病病原学检测方法。在使用中各自又有其特点。

     

    Abstract: Three methods were evaluated:(1)an assay of amplification for a 245 bp segment derived from therepetitive DNA IS986 specific for mycobacterium tuberculosis cornplex was developed by using polymerasechain reaction(PCR),(2)a sandwiched enzvme linked imnlunosorbant assay(ELISA)was developed by usingmonoclonal antibody(Mab)TB15-C3 that recognized specific antigenic determinant of mycobacterium tuber-culosis, and (3)a conventional acid-fast stain,Ten strains of acid-fast mvcobacteria and 2 strains of non-my-cobacteria were tested.The specific PCR product was obtained only from mycobacterium tuberculosis com-plex.Tests for M,tuberculosis and BCG,M.avium and M.scrofulaceum were positive by Mab-ELISA anal-vsis.All strains of acid-fast mycobacteria were positive by conventional acid-fast stain.The sensitivity of PCR in the detection of M. tuberculosis genomic DNA and bacteria suspension was lpg,corresponding to 100 viable bacteria cells.The sensitivity of detection of PPD by Mab-ELISA was 15ng/ml,and the sensitivity of conventional acid-fast stain was 10 ̄4 viable bacteria cells.One hundred and thir-ty-eight clinical specimens from patients with tuberculosis were examined by the three methods.The positivi-ty rate of PCR was higher than that of conventional acid-fast stain(P<0.05).The positivity rate of Mab-ELISA in detection of specific antigenic determinant was much higher than that of both conventional acid-faststain and PCR(P<0.01).The false positivity rate of Mab-ELISA was not remarkably higher than that ofPCR (P>0.05).Our results suggest that PCR and Mab-ELISA appear to be specific,sensitive and rapidtechniques for the diagnosis of tuberculosis.

     

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