title: Identification of Antibody Targets for Tuberculosis Serology using High-Density Nucleic Acid Programmable Protein Arrays.
abstract: Better and more diverse biomarkers for the development of simple point-of-care tests for active tuberculosis (TB), a clinically heterogeneous disease, are urgently needed. We generated a proteomicMycobacterium tuberculosis(Mtb) High-Density Nucleic Acid Programmable Protein Array (HD-NAPPA) that used a novel multiplexed strategy for expedited high-throughput screening for antibody responses to theMtbproteome. We screened sera from HIV uninfected and coinfected TB patients and controls (n= 120) from the US and South Africa (SA) using the multiplex HD-NAPPA for discovery, followed by deconvolution and validation through single protein HD-NAPPA with biologically independent samples (n= 124). We verified the top proteins with enzyme-linked immunosorbent assays (ELISA) using the original screening and validation samples (n= 244) and heretofore untested samples (n= 41). We identified 8 proteins with TB biomarker value; four (Rv0054, Rv0831c, Rv2031c and Rv0222) of these were previously identified in serology studies, and four (Rv0948c, Rv2853, Rv3405c, Rv3544c) were not known to elicit antibody responses. Using ELISA data, we created classifiers that could discriminate patients' TB status according to geography (US or SA) and HIV (HIV- or HIV+) status. With ROC curve analysis under cross validation, the classifiers performed with an AUC for US/HIV- at 0.807; US/HIV+ at 0.782; SA/HIV- at 0.868; and SA/HIV+ at 0.723. With this study we demonstrate a new platform for biomarker/antibody screening and delineate its utility to identify previously unknown immunoreactive proteins.
authors.affiliation: From the ‡The Virginia G Piper Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University, Tempe, Arizona, 85287.
authors.forename: Lusheng
authors.initials: L
authors.lastname: Song
authors.affiliation: From the ‡The Virginia G Piper Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University, Tempe, Arizona, 85287.
authors.forename: Garrick
authors.initials: G
authors.lastname: Wallstrom
authors.affiliation: §State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences-Beijing (PHOENIX Center), Beijing Institute of Radiation Medicine, Beijing, 102206, China.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Yu
authors.affiliation: From the ‡The Virginia G Piper Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University, Tempe, Arizona, 85287.
authors.forename: Marika
authors.initials: M
authors.lastname: Hopper
authors.affiliation: From the ‡The Virginia G Piper Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University, Tempe, Arizona, 85287.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Van Duine
authors.affiliation: From the ‡The Virginia G Piper Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University, Tempe, Arizona, 85287.
authors.forename: Jason
authors.initials: J
authors.lastname: Steel
authors.affiliation: From the ‡The Virginia G Piper Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University, Tempe, Arizona, 85287.
authors.forename: Jin
authors.initials: J
authors.lastname: Park
authors.affiliation: From the ‡The Virginia G Piper Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University, Tempe, Arizona, 85287.|¶Engineering Arts LLC, Tempe, Arizona 85287.
authors.forename: Peter
authors.initials: P
authors.lastname: Wiktor
authors.affiliation: ¶Engineering Arts LLC, Tempe, Arizona 85287.
authors.forename: Peter
authors.initials: P
authors.lastname: Kahn
authors.affiliation: ¶Engineering Arts LLC, Tempe, Arizona 85287.
authors.forename: Al
authors.initials: A
authors.lastname: Brunner
authors.affiliation: ‖Department of Internal Medicine, Edendale Hospital, University of KwaZulu-Natal, Pietermaritzburg, South Africa.
authors.forename: Douglas
authors.initials: D
authors.lastname: Wilson
authors.affiliation: **Departments of Medicine and.
authors.forename: Elizabeth R
authors.initials: ER
authors.lastname: Jenny-Avital
authors.affiliation: From the ‡The Virginia G Piper Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University, Tempe, Arizona, 85287.
authors.forename: Ji
authors.initials: J
authors.lastname: Qiu
authors.affiliation: From the ‡The Virginia G Piper Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University, Tempe, Arizona, 85287.
authors.forename: Joshua
authors.initials: J
authors.lastname: Labaer
authors.affiliation: From the ‡The Virginia G Piper Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University, Tempe, Arizona, 85287; jacqueline.achkar@einstein.yu.edu mitch.magee@asu.edu.
authors.forename: D Mitchell
authors.initials: DM
authors.lastname: Magee
authors.affiliation: **Departments of Medicine and jacqueline.achkar@einstein.yu.edu mitch.magee@asu.edu.|‡‡Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York 10461.
authors.forename: Jacqueline M
authors.initials: JM
authors.lastname: Achkar
mesh_terms: D000328:Adult; D000368:Aged; D001426:Bacterial Proteins / Q000276:immunology*; D015415:Biomarkers / Q000097:blood; D060085:Coinfection; D005260:Female; D015658:HIV Infections / Q000097:blood*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009169:Mycobacterium tuberculosis / Q000276:immunology / Q000378:metabolism*; D040081:Protein Array Analysis / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D012372:ROC Curve; D058248:Serum Bactericidal Antibody Assay / Q000379:methods*; D013019:South Africa; D014376:Tuberculosis / Q000276:immunology*; D014481:United States; D055815:Young Adult
chemical_list: D001426:Bacterial Proteins; D015415:Biomarkers