A Peptidomic Approach to Identify Novel Antigen Biomarkers for the Diagnosis of Tuberculosis

Hongmei Chen; Shanshan Li; Weijie Zhao; Jiaheng Deng; Zhuohong Yan; Tingting Zhang; Shu' An Wen; Haiping Guo; Lei Li; Jianfeng Yuan; Hongtao Zhang; Liping Ma; Jianhua Zheng; Mengqiu Gao; Yu Pang

doi:10.2147/IDR.S373652

A Peptidomic Approach to Identify Novel Antigen Biomarkers for the Diagnosis of Tuberculosis

Infect Drug Resist. 2022 Aug 18:15:4617-4626. doi: 10.2147/IDR.S373652. eCollection 2022.

Authors

Hongmei Chen^#¹, Shanshan Li^#², Weijie Zhao^#³, Jiaheng Deng^#⁴, Zhuohong Yan⁵, Tingting Zhang², Shu' An Wen², Haiping Guo², Lei Li⁶, Jianfeng Yuan⁶, Hongtao Zhang⁵, Liping Ma¹, Jianhua Zheng^#⁴, Mengqiu Gao¹, Yu Pang²

Affiliations

¹ Department of Tuberculosis, Beijing Chest Hospital, Capital Medical University/Beijing Tuberculosis & Thoracic Tumor Research Institute, Beijing, People's Republic of China.
² Department of Bacteriology and Immunology, Beijing Chest Hospital, Capital Medical University/Beijing Tuberculosis & Thoracic Tumor Research Institute, Beijing, People's Republic of China.
³ Clinical Trial Agency Office, Beijing Chest Hospital, Capital Medical University/Beijing Tuberculosis & Thoracic Tumor Research Institute, Beijing, People's Republic of China.
⁴ Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, and Center for Tuberculosis Research, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, People's Republic of China.
⁵ Department of Central Laboratory, Beijing Chest Hospital, Capital Medical University/Beijing Tuberculosis & Thoracic Tumor Research Institute, Beijing, People's Republic of China.
⁶ Electral Safety Research & Development Center, Beijing Normal University, Zhuhai, People's Republic of China.

^# Contributed equally.

Abstract

Background: Here, we conducted a peptidomic study in murine model to identify novel antigen biomarkers for the diagnosis of tuberculosis (TB) with improved performance.

Methods: Four recombinant proteins, including Mycobacterium tuberculosis protein 32 (MPT32), Mycobacterium tuberculosis protein 64 (MPT64), culture filtrate protein 10 (CFP10), and phosphate ABC transporter substrate-binding lipoprotein (PstS1) were expressed and intravenously injected into BALB/c mice. The serum were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The concentrations of candidate peptides in serum of suspected TB patients were determined using competitive enzyme-linked immunosorbent assay.

Results: A total of 65 peptides from 4 MTB precursor recombinant proteins were identified in mouse serum by LC-MS/MS, of which 5 peptides were selected as candidates for serological analysis. The concentrations of peptides MPT64-2, CFP10-2 and PstS1-2 in TB patients were significantly higher than those in non-TB patients. MPT64-2 exhibited the most promising sensitivity (81.4%), followed by PstS1-2 and CFP10-2. In addition, PstS1-2 had the highest specificity (93.3%), followed by CFP10-2 and MPT64-2. According to the area under the curve (AUC), MPT64-2 (AUC = 0.863), PstS1-2 (AUC = 0.812) and CFP10-2 (AUC = 0.809) exhibited better diagnostic validity.

Conclusion: We develop an effective approach to identify new antigen biomarkers via LC-MS/MS-based peptidomics. Multiple peptides exhibit promising efficacy in diagnosis of active TB patients.

Keywords: Mycobacterium tuberculosis protein 64; diagnostics; enzyme-linked immunosorbent assay; peptidomics; tuberculosis.

Grants and funding

This study was supported by the Beijing Hospitals Authority Ascent Plan (DFL20191601), the Capital’s Funds for Health Improvement and Research (2020-1-1041), the Beijing Hospitals Authority Clinical Medicine Development of Special Funding (ZYLX202122), CAMS Innovation Fund for Medical Sciences (2021-I2M-1-037) and the National Science and Technology Major Project of China (2017ZX10201301-002-003). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of manuscript.