Department of Central Laboratory.
Department of Beijing Key Laboratory for Drug Resistance Tuberculosis Research, Beijing Chest Hospital, Capital Medical University, Beijing Tuberculosis and Thoracic Tumour Research Institute, Beijing.
Int J Tuberc Lung Dis. 2018 Aug 1;22(8):959-965. doi: 10.5588/ijtld.18.0060.
To evaluate antibody responses to panels of Mycobacterium tuberculosis antigens for serological diagnosis of active tuberculosis (TB).
We cloned, expressed and purified 10 M. tuberculosis recombinant proteins 38KD, MPT32 (M. tuberculosis protein 32), MPT64, EspC (ESX-1 secretion-associated protein), Mtb81, Rv3881, Rv3425, Rv0222, Rv3872 and CFP21 (culture filtrate protein 21), and obtained lipoarabinomannan (LAM) polysaccharide antigen from BEI Resources. The plasma immunoglobulin (Ig)G titre responses to the 11 antigens based on 45 patients with pulmonary TB (PTB) and 30 healthy controls (HCs) were first evaluated using enzyme-linked immunosorbent assays. Antigens with high sensitivities were then selected for further investigation in 200 PTB patients (121 smear- or culture-positive patients, 79 smear- or culture-negative patients) and 152 HCs.
LAM, 38KD, MPT32, MPT64, EspC and Mtb81 were chosen. The LAM, 38KD, MPT32 and EspC IgG titres were significantly higher in Bacterium-negative TB patients than in HCs, except for MPT64 and Mtb81. The sensitivity of the individual antigens for detecting antibodies ranged from 21.5% to 67.0%, with 74.3-98.0% specificity. The sensitivity of MPT32 was higher than that of 38KD at a high level of specificity. The six-antigen combination reached a sensitivity of 69.6% in bacterium-negative TB patients, with 77.0% specificity.
The combination panel had markedly improved sensitivity, but specificity requires further enhancement.
评估针对结核分枝杆菌抗原的抗体反应，以用于活动性肺结核（TB）的血清学诊断。
我们克隆、表达和纯化了 10 种结核分枝杆菌重组蛋白 38KD、MPT32（结核分枝杆菌蛋白 32）、MPT64、EspC（ESX-1 分泌相关蛋白）、Mtb81、Rv3881、Rv3425、Rv0222、Rv3872 和 CFP21（培养滤液蛋白 21），并从 BEI 资源中获得了脂阿拉伯甘露聚糖（LAM）多糖抗原。首先使用酶联免疫吸附试验评估了 45 例肺结核（PTB）患者和 30 例健康对照（HC）患者对基于 11 种抗原的血浆免疫球蛋白（IgG）滴度反应。然后选择高灵敏度的抗原，在 200 例 PTB 患者（121 例涂片或培养阳性患者，79 例涂片或培养阴性患者）和 152 例 HC 中进行进一步研究。
选择了 LAM、38KD、MPT32、MPT64、EspC 和 Mtb81。LAM、38KD、MPT32 和 EspC 的 IgG 滴度在细菌阴性的 TB 患者中明显高于 HC，除了 MPT64 和 Mtb81。各抗原检测抗体的灵敏度范围为 21.5%至 67.0%，特异性为 74.3-98.0%。MPT32 的灵敏度高于 38KD，特异性较高。在细菌阴性的 TB 患者中，六抗原组合的灵敏度达到 69.6%，特异性为 77.0%。
组合面板的敏感性有了明显提高，但特异性需要进一步提高。
