Wadee A A, Boting L, Reddy S G
Department of Immunology of the South African Institute for Medical Research, Johannesburg.
J Clin Microbiol. 1990 Dec;28(12):2786-91. doi: 10.1128/jcm.28.12.2786-2791.1990.
This study describes the development of an enzyme-linked immunosorbent assay (ELISA) to detect Mycobacterium tuberculosis antigens in body fluids. A double-antibody sandwich procedure that used human and rabbit anti-M. tuberculosis immunoglobulin G antibodies was followed. The ELISA was able to detect as little as 0.8 micrograms of protein of M. tuberculosis sonic extract. Of 253 cerebrospinal fluid specimens submitted for analysis, 11 (4.3%) false-positive results were recorded. Analysis of 317 pleural and ascitic fluid specimens resulted in 6 (1.9%) false-positive recordings. No false-negative results were recorded for any of the body fluids tested. This technique is rapid (5.5 h) and sensitive, may be developed and used in many laboratories with limited resources, and may prove useful in the diagnosis of extrapulmonary and pulmonary tuberculoses. Analysis of these body fluids by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western immunoblotting indicated that the antibody used in the ELISA detects a mycobacterial antigen of 43 kDa. Such antigens were not detected in body fluids of nontuberculous patients.
本研究描述了一种用于检测体液中结核分枝杆菌抗原的酶联免疫吸附测定（ELISA）的开发。采用了一种双抗体夹心程序，该程序使用了人和兔抗结核分枝杆菌免疫球蛋白G抗体。该ELISA能够检测低至0.8微克的结核分枝杆菌超声提取物蛋白。在提交分析的253份脑脊液标本中，记录到11份（4.3%）假阳性结果。对317份胸水和腹水标本的分析产生了6份（1.9%）假阳性记录。在所检测的任何体液中均未记录到假阴性结果。该技术快速（5.5小时）且灵敏，可在许多资源有限的实验室中开发和使用，并且可能在肺外和肺结核的诊断中证明有用。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和蛋白质免疫印迹对这些体液进行分析表明，ELISA中使用的抗体检测到一种43 kDa的分枝杆菌抗原。在非结核患者的体液中未检测到此类抗原。
