Preprint / Version 1

Enrichment-assisted PhIP-seq discovers a serum antibody peptide signature for active tuberculosis

This article is a preprint and has not been certified by peer review.

Authors

    Haoyun Xu,  
    Haoyun Xu
    Mingxiang Huang,  
    Mingxiang Huang
    Zhaowei Xu,  
    Zhaowei Xu
    Jiaoxiang Wu,  
    Jiaoxiang Wu
    Chongxiang Tong,  
    Chongxiang Tong
    Yunhong Tan,  
    Yunhong Tan
    • Hunan Institute for Tuberculosis Control
    Quan Wang,  
    Quan Wang
    • Sixth People's Hospital of Xinjiang Uygur Autonomous Region
    Xundi Bao,  
    Xundi Bao
    Sheng Tan,  
    Sheng Tan
    Shujuan Guo,  
    Shujuan Guo
    Zigan Sha,  
    Zigan Sha
    Jiahao Xie,  
    Jiahao Xie
    Hongzhen Di,  
    Hongzhen Di
    Mingliang Ma,  
    Mingliang Ma
    • AbCode Biotechnology Co., Ltd
    Jinhua Wu,  
    Jinhua Wu
    • AbCode Biotechnology Co., Ltd
    Yang Li,  
    Yang Li
    • AbCode Biotechnology Co., Ltd
    Huan Qi,  
    Huan Qi
    Sheng-ce Tao
    Sheng-ce Tao
Categories
Immunology & Microbiology
Keywords
Tuberculosis; Mycobacterium tuberculosis; H37Rv; Biomarker; PhIP-seq

Abstract

Serological diagnosis of active tuberculosis has been limited by inconsistent performance of historical antibody tests, partly reflecting incomplete and candidate-driven antigen discovery. Here, we developed enrichment-assisted phage immunoprecipitation sequencing (ePhIP-seq) to systematically identify serum IgG-reactive peptide biomarkers for active tuberculosis. We constructed a proteome-wide Mycobacterium tuberculosis H37Rv T7 phage-display library containing 86,287 overlapping 30-amino-acid peptides and enriched H37Rv-reactive serum antibodies before PhIP-seq screening. In a discovery cohort of 30 patients with active tuberculosis and 30 healthy controls, ePhIP-seq identified 277 differential peptide candidates, from which 21 were selected for orthogonal peptide microarray validation. 15 peptides showed higher IgG reactivity in active tuberculosis and were used for model development. A compact 3-marker candidate signature, Model C359, comprising MTB-N-9, MTB-N-10, and MTB-N-14, achieved an apparent AUC of 0.951 and a bootstrap optimism-corrected AUC of 0.942 in the model-building cohort. In a held-out internal test set, Model C359 achieved an AUC of 0.886 for active tuberculosis versus healthy controls. In exploratory disease-control analysis against pulmonary Mycobacterium intracellulare infection, the AUC was 0.672. These findings nominate a compact serum antibody peptide signature for further validation and support ePhIP-seq as a reusable antigen-discovery strategy for diseases in which pathogen-specific antibody signals may be weak or diluted within total serum IgG.

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2026-09-15

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Xu, H., Huang, M., Xu, Z., Wu, J., Tong, C., Tan, Y., Wang, Q., Bao, X., Tan, S., Guo, S., Sha, Z., Xie, J., Di, H., Ma, M., Wu, J., Li, Y., Qi, H., & Tao, S.- ce. (2026). Enrichment-assisted PhIP-seq discovers a serum antibody peptide signature for active tuberculosis. LangTaoSha Preprint Server. https://doi.org/10.65215/LTSpreprints.2026.09.14.000337

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Declaration of Competing Interests

Details of all competing interests to be disclosed are as follows:

Sheng-ce Tao is a co-founder of AbCode Biotechnology Co., Ltd. Sheng-ce Tao, Huan Qi, Shujuan Guo and Haoyun Xu are co-inventors on a pending patent application (202510411480.9) related to TbMap construction described in this manuscript. The other authors declare no competing interests.

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