Identification of Antibody Targets for Tuberculosis Serology using High-Density Nucleic Acid Programmable Protein...

Area of Science:

• Immunology
• Proteomics
• Infectious Diseases

Background:

• Active tuberculosis (TB) is a heterogeneous disease requiring improved diagnostic biomarkers.
• Current diagnostic methods for TB lack diversity and simplicity for point-of-care applications.
• Need for novel serological biomarkers to detect active TB, especially in diverse populations.

Purpose of the Study:

• To develop and utilize a novel proteomic array for high-throughput screening of antibody responses to Mycobacterium tuberculosis (Mtb) antigens.
• To identify novel TB biomarkers for improved point-of-care diagnostic tests.
• To evaluate the performance of identified biomarkers in classifying TB status across different geographical locations and HIV statuses.

Main Methods:

• Generation of a Mycobacterium tuberculosis (Mtb) High-Density Nucleic Acid Programmable Protein Array (HD-NAPPA) using a multiplexed strategy.
• Screening of sera from TB patients and controls (HIV-uninfected and coinfected) from the US and South Africa (SA) using multiplex and single-protein HD-NAPPA.
• Validation of identified protein biomarkers using enzyme-linked immunosorbent assays (ELISA) and development of diagnostic classifiers.

Main Results:

• Identification of 8 proteins with TB biomarker value, including 4 novel immunoreactive proteins (Rv0948c, Rv2853, Rv3405c, Rv3544c).
• Development of diagnostic classifiers with significant Area Under the Curve (AUC) values for discriminating TB status based on geography and HIV status (e.g., US/HIV-: 0.807, SA/HIV-: 0.868).
• Demonstration of a new platform for biomarker discovery and validation of antibody responses.

Conclusions:

• The novel HD-NAPPA platform is effective for high-throughput screening and discovery of Mtb immunoreactive proteins.
• The identified biomarkers show potential for developing simple, point-of-care diagnostic tests for active TB.
• The study highlights the utility of proteomic arrays in identifying novel biomarkers for complex infectious diseases like TB.