CFP10 and ESAT6 aptamers as effective Mycobacterial antigen diagnostic reagents
CFP10 and ESAT6 aptamers as effective Mycobacterial antigen diagnostic reagents
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CFP10和ESAT6适体作为有效的分枝杆菌抗原诊断试剂。
DOI：
10.1016/j.jinf.2014.05.015
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发表时间：
2014-12-01
影响因子：
28.2
通讯作者：
Zhang, Xiao-Lian
中科院分区：
文献类型：
--
作者：
Tang, Xiao-Lei;Zhou, Ya-Xiong;Zhang, Xiao-Lian
有效的分枝杆菌抗原诊断试剂的开发仍然是一个高度优先事项。6-kDa早期分泌的抗原靶标（ESAT 6）和10-kDa培养滤液蛋白（CFP 10）由毒性结核分枝杆菌（M. tb），并且不存在于非毒性卡介苗（BCG）中。在这项研究中，我们使用了指数富集配体系统进化（SELEX）技术来筛选特异性结合ESAT 6-CFP 10（CE）蛋白的功能性ssDNA适体“抗体”。选择的单一ssDNA适体（CE 24和CE 15）对CFP 10（CE 24：K-d = 3.75 x 10（-7）M）和ESAT 6（CE 15：K-d = 1.6 x 10（-7）M）表现出最高的特异性和结合亲和力。我们进一步用酶联寡核苷酸分析法（ELISA）检测了活动性肺结核（TB）患者、肺外TB患者和健康献血员血清中CFP 10和ESAT 6蛋白。结果表明，敏感性和特异性分别为100%和94.1%（使用基于CE 24适体的cDNA）和89.6%和94.1%（使用基于CE 15适体的cDNA）。在基于适体的ELISA和T-SPOT TB测定之间观察到良好的相关性。因此，我们的研究表明，CE 24和CE 15作为早期抗原诊断剂不仅对活动性肺结核、肺外结核，而且可能对潜伏性结核感染和免疫缺陷型结核具有潜在的广泛应用。(C)2014年，英国传染病协会。由爱思唯尔有限公司出版。保留所有权利。
The development of effective Mycobacterial antigen diagnostic reagents remains a high priority. The 6-kDa early secreted antigenic target (ESAT6) and 10-kDa culture filtrate protein (CFP10) are secreted early by virulent Mycobacterium tuberculosis (M. tb) and are not present in the non-virulent Bacillus Calmette-Guerin (BCG). In this study, we used a Systematic Evolution of Ligands by Exponential Enrichment (SELEX) technique to screen for a functional ssDNA aptamer "antibody" that specifically bound to ESAT6-CFP10 (CE) protein. The selected single ssDNA aptamers (CE24 and CE15) demonstrated the highest specificity and binding affinity to CFP10 (CE24: K-d = 3.75 x 10(-7) M) and ESAT6 (CE15: K-d = 1.6 x 10(-7) M). We further detected CFP10 and ESAT6 proteins in serum samples from active pulmonary tuberculosis (TB) patients, extrapulmonary TB patients and healthy donors by using an enzyme-linked oligonucleotide assay (ELONA). The results showed that the sensitivity and specificity were 100% and 94.1% (using CE24 aptamer-based ELONA) and 89.6% and 94.1% (using CE15 aptamer-based ELONA), respectively. A good correlation was observed between aptamer-based ELONA and T-SPOT TB assay. Thus, our study suggests that CE24 and CE15 have potentially broad applications as early antigen diagnostic agents not only for active pulmonary TB, extrapulmonary TB, but also possibly for latent TB infection and TB with immune-deficiency. (C) 2014 The British Infection Association. Published by Elsevier Ltd. All rights reserved.