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通过蛋白质印迹法鉴定结核性脑膜炎患者脑脊液中针对结核分枝杆菌抗原的抗体反应。

Identification of antibody responses to Mycobacterium tuberculosis antigens in the CSF of tuberculous meningitis patients by Western blotting.

作者信息

Patil S A, Gourie-Devi M, Chaudhuri J R, Chandramuki A

机构信息

Department of Microbiology, National Institute of Mental Health and Neurosciences, Bangalore, India.

出版信息

Clin Immunol Immunopathol. 1996 Oct;81(1):35-40. doi: 10.1006/clin.1996.0154.

Abstract

One of the adjunctive modes of diagnosing tuberculous meningitis (TBM) is to detect immune responses in the cerebrospinal fluid (CSF) to the Mycobacterium tuberculosis antigen. Up to 70% of clinical TBM reveal the presence of antimycobacterial antibody by the enzyme-linked immunosorbant assay. Defining the specificity of this immune response by Western blotting on separated M. tuberculosis antigen has been attempted in this study. Only antimycobacterial antibody-positive TBM cases were included in the study. An analysis of 30 such TBM cases showed a major immune reactivity to the 30- to 40-kDa region (93%) while a lower degree of immune reactivity was seen to the 14-kDa region (87%) and to the 18- to 25-kDa region (60%). Grossly the antibody reactivity on Western blot correlated with the ELISA results. Assessment of antimycobacterial antibody in the neurologic control CSF samples of pyogenic meningitis [n = 10], cryptococcal meningitis [6], neurocysticercosis [28], neurosyphilis [8], viral meningoencephalitis [8], carcinomatous meningitis [8], iatrogenic meningitis [6], and nonneurological control CSF samples from patients undergoing spinal anesthesia [20] revealed the presence of antibody in the CSF of 2 of the 10 pyogenic meningitis and 5 of the 28 neurocysticercosis cases. A Western blot analysis of these 7 cases revealed immune reactivity to 30- to 40-kDa regions only in 2 cases (1 of pyogenic and 1 of neurocysticercosis). The remaining 5 CSF samples did not reveal any immune reactivity on Western blotting, although ELISA demonstrated antimycobacterial antibodies. The antibody response to M. tuberculosis lipoarabinomannan and 38-kDa antigen by ELISA revealed 70.58 and 41.17% positivity, respectively. Thus this study has demonstrated that, by Western blotting, the major immune response is to the 30- to 40-kDa region, namely, lipoarabinomannan. Further, this finding will be useful for specific immunodiagnosis of the TBM.

摘要

诊断结核性脑膜炎(TBM)的辅助方法之一是检测脑脊液(CSF)中针对结核分枝杆菌抗原的免疫反应。高达70%的临床TBM病例通过酶联免疫吸附测定法显示存在抗分枝杆菌抗体。本研究尝试通过对分离的结核分枝杆菌抗原进行蛋白质印迹法来确定这种免疫反应的特异性。该研究仅纳入了抗分枝杆菌抗体阳性的TBM病例。对30例此类TBM病例的分析显示,主要免疫反应针对30至40 kDa区域(93%),而对14 kDa区域(87%)和18至25 kDa区域(60%)的免疫反应程度较低。总体而言,蛋白质印迹法上的抗体反应性与酶联免疫吸附测定结果相关。对化脓性脑膜炎[10例]、隐球菌性脑膜炎[6例]、神经囊尾蚴病[28例]、神经梅毒[8例]、病毒性脑膜脑炎[8例]、癌性脑膜炎[8例]、医源性脑膜炎[6例]的神经对照CSF样本以及接受脊髓麻醉患者的非神经对照CSF样本[20例]中的抗分枝杆菌抗体进行评估,结果显示10例化脓性脑膜炎病例中有2例以及28例神经囊尾蚴病病例中有5例的CSF中存在抗体。对这7例病例进行蛋白质印迹分析发现,只有2例(1例化脓性脑膜炎和1例神经囊尾蚴病)对30至40 kDa区域有免疫反应。其余5份CSF样本在蛋白质印迹法上未显示任何免疫反应,尽管酶联免疫吸附测定显示存在抗分枝杆菌抗体。通过酶联免疫吸附测定法对结核分枝杆菌脂阿拉伯甘露聚糖和38 kDa抗原的抗体反应分别显示阳性率为70.58%和41.17%。因此,本研究表明,通过蛋白质印迹法,主要免疫反应针对30至40 kDa区域,即脂阿拉伯甘露聚糖。此外,这一发现将有助于TBM的特异性免疫诊断。

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