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探索一些新的分泌蛋白,用于结核分枝杆菌的伴随诊断。

Exploration of some new secretory proteins to be employed for companion diagnosis of Mycobacterium tuberculosis.

机构信息

Department of Microbiology, CSIR-Central Drug Research Institute, Lucknow, 226031, India.

Department of Respiratory Medicine, KGMU, UP, Lucknow, 226003, India.

出版信息

Immunol Lett. 2019 May;209:67-74. doi: 10.1016/j.imlet.2019.03.010. Epub 2019 Mar 18.

Abstract

Tuberculosis (TB) is a highly infectious disease and its early and precise diagnosis is essential to reduce morbidity and mortality of patients. Since the routine diagnostic tests (like Monteux, AFB smear microscopy, chest X-Ray) do not give infallible results, additional tests are always recommended. Therefore to address the concerns about non-specificity of the present battery of diagnostic tests, we have attempted to analyze some unique secretory antigens which could be able to identify the stage specific infection of MTB. In this study, we have used recombinant proteins CFP-10, ESAT-6, Ag85 A, Ag85B, Ag85C, PE3, PE4 and Mycp1 to eliminate heterogeneity and cross reactivity in clinical diagnosis. Amplified genes were cloned and over-expressed in Escherichia coli BL21 (DE3). The recombinantly purified proteins were used as antigens against 158 sera samples of TB patients. Secretory proteins showed better response than the PPD control. Among all the used antigens PE3 and PE4 proteins showed better reactivity levels among all the groups of TB patients. The secretions of CFP-10 and ESAT-6 were also higher as compared to other secretory proteins like Ag85 A, Ag85B, Ag85C and MycP1.The clinical use of these newly identified secretory antigens could be of significant value for the confirmatory, rapid, simple and low-cost diagnosis of TB patients.

摘要

结核病(TB)是一种高度传染性疾病,早期和准确的诊断对于降低患者的发病率和死亡率至关重要。由于常规诊断测试(如蒙特氏试验、AFB 涂片显微镜检查、胸部 X 光检查)并不能给出可靠的结果,因此总是建议进行额外的测试。因此,为了解决目前诊断测试特异性不足的问题,我们试图分析一些独特的分泌性抗原,这些抗原能够识别 MTB 的特定感染阶段。在这项研究中,我们使用了重组蛋白 CFP-10、ESAT-6、Ag85A、Ag85B、Ag85C、PE3、PE4 和 Mycp1 来消除临床诊断中的异质性和交叉反应性。扩增的基因被克隆并在大肠杆菌 BL21(DE3)中过表达。重组纯化的蛋白被用作针对 158 份结核病患者血清样本的抗原。分泌蛋白比 PPD 对照显示出更好的反应性。在所有使用的抗原中,PE3 和 PE4 蛋白在所有结核病患者组中表现出更好的反应水平。与其他分泌蛋白(如 Ag85A、Ag85B、Ag85C 和 MycP1)相比,CFP-10 和 ESAT-6 的分泌量也更高。这些新鉴定的分泌性抗原的临床应用对于结核病患者的确认、快速、简单和低成本诊断可能具有重要价值。

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