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联合使用蛋白质印迹法/酶联免疫吸附测定法以改进人类结核病的血清学诊断。

Combined use of Western blot/ELISA to improve the serological diagnosis of human tuberculosis.

作者信息

Beck Sandra Trevisan, Leite O M, Arruda R S, Ferreira A W

机构信息

Federal University of Santa Maria, Santa Maria, RS, Brazil.

出版信息

Braz J Infect Dis. 2005 Feb;9(1):35-43. doi: 10.1590/s1413-86702005000100007. Epub 2005 Jun 6.

Abstract

Two recombinant antigens and a crude bacterial antigen of a wild M. tuberculosis strain were used to detect specific IgG antibodies in sera from 52 patients with pulmonary tuberculosis, confirmed by an acid-fast smear and serum culture of these patients and that of 25 contacts. The patients were not infected with HIV. We evaluated the sensitivity and specificity of ELISA, based on the recombinant TbF6 and TbF6/DPEP antigen and a search for reactivity patterns in the Western blot technique, using whole mycobacterium antigen. Serum samples from 22 healthy individuals and from 30 patients with lung diseases other than tuberculosis were used as controls. The best ELISA results were obtained with the TbF6/DPEP antigen combination, which gave 85% sensitivity and 91% specificity. ELISA sensitivity improved from 85% to 92% when the Western blot results were used. Western blot specificity was 100% when antibody reactivity with different antigenic bands was analyzed and associated. The association of TbF6/DPEP antigens used in ELISA with specific patterns of reactivity determined by Western blot can help make an identification when classic methods for the diagnosis of pulmonary tuberculosis are not sufficient.

摘要

两种重组抗原和一株野生结核分枝杆菌菌株的粗细菌抗原,用于检测52例经抗酸涂片和血清培养确诊为肺结核患者以及25名接触者血清中的特异性IgG抗体。这些患者未感染艾滋病毒。我们基于重组TbF6和TbF6/DPEP抗原评估了ELISA的敏感性和特异性,并利用全分枝杆菌抗原在蛋白质印迹技术中寻找反应模式。来自22名健康个体和30例非结核性肺部疾病患者的血清样本用作对照。使用TbF6/DPEP抗原组合获得了最佳ELISA结果,其敏感性为85%,特异性为91%。当采用蛋白质印迹结果时,ELISA敏感性从85%提高到92%。分析并关联抗体与不同抗原条带的反应性时,蛋白质印迹特异性为100%。ELISA中使用的TbF6/DPEP抗原与蛋白质印迹确定的特定反应模式相结合,有助于在肺结核经典诊断方法不足时进行鉴别诊断。

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