Yang Hua, Sha Wei, Song Peng, Liu Zhonghua, Qin Lianhua, Huang Xiaocheng, Lu Junmei, Wang Jie, Duthie Malcolm S, Xiao Heping, Hu Zhongyi
Shanghai Key Laboratory of Tuberculosis, Clinic and Research Center of Tuberculosis, Shanghai Pulmonary Hospital, Tongji University School of Medicine, 507 Zhengmin Road, Shanghai, 200433, People's Republic of China.
Department of Microbiology and Immunology, Tongji University School of Medicine, 1239 Siping Road, Shanghai, 200433, People's Republic of China.
Appl Microbiol Biotechnol. 2016 Mar;100(5):2279-87. doi: 10.1007/s00253-015-7122-z. Epub 2015 Nov 18.
Although serological detection is a practical strategy for early detection and diagnosis of tuberculosis (TB), inconsistent and imprecise estimates of sensitivity and specificity block its development and application for clinic. New or alternative serological antigens with improved accuracy are urgently needed. A phage-displayed random peptide library was employed to screen for immunoactive peptides using specific immunoglobulin G (IgG) of TB patients as target molecules. With two screening strategies, 20 single phages displaying different sequences were obtained and no sequence homology was found among these phages. From the results of phage-ELISA, H12, TB6, TB15, and TB18 phages showed higher affinity to IgGs from TB patients(S/N ≥2.1) and were identified as the positive clones. Significant differences in the detection values of sera from 47 TB patients and 37 healthy individuals were found for these four phage clones. According to the reactivity of 284 human sera to synthetic H12, TB6, TB15, and TB18 peptides as determined by ELISA, TB15 showed significantly higher areas under the curve (AUC) and sensitivity than other peptides, providing a lead molecule for the development of new serology diagnostic strategies for TB.
虽然血清学检测是结核病（TB）早期检测和诊断的一种实用策略，但敏感性和特异性的估计不一致且不准确阻碍了其在临床上的发展和应用。迫切需要具有更高准确性的新型或替代性血清学抗原。利用噬菌体展示随机肽库，以结核病患者的特异性免疫球蛋白G（IgG）作为靶分子筛选免疫活性肽。通过两种筛选策略，获得了20个展示不同序列的单个噬菌体，这些噬菌体之间未发现序列同源性。从噬菌体ELISA结果来看，H12、TB6、TB15和TB18噬菌体对结核病患者的IgG显示出更高的亲和力（S/N≥2.1），并被鉴定为阳性克隆。发现这四个噬菌体克隆对47例结核病患者和37例健康个体血清的检测值存在显著差异。根据ELISA测定的284份人血清对合成的H12、TB6、TB15和TB18肽的反应性，TB15显示出比其他肽更高的曲线下面积（AUC）和敏感性，为开发新的结核病血清学诊断策略提供了一个先导分子。
