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Title: Identification of circulating bacterial antigens by in vivo microbial antigen discovery
Authors: Dana E. Nuti
Reva B. Crump
Farida Dwi Handayani
Narisara Chantratita
Sharon J. Peacock
Richard Bowen
Philip L. Felgner
D. Huw Davies
Terry Wu
C. Rick Lyons
Paul J. Brett
Mary N. Burtnick
Thomas R. Kozel
David P. AuCoina
University of Nevada School of Medicine
Mahidol University
University of Cambridge
Colorado State University
University of California, Irvine
University of New Mexico Health Sciences Center
University of South Alabama
Keywords: Immunology and Microbiology
Issue Date: 1-Jul-2011
Citation: mBio. Vol.2, No.4 (2011)
Abstract: Detection of microbial antigens in clinical samples can lead to rapid diagnosis of an infection and administration of appropriate therapeutics. A major barrier in diagnostics development is determining which of the potentially hundreds or thousands of antigens produced by a microbe are actually present in patient samples in detectable amounts against a background of innumerable host proteins. In this report, we describe a strategy, termed in vivo microbial antigen discovery (InMAD), that we used to identify circulating bacterial antigens. This technique starts with "InMAD serum," which is filtered serum that has been harvested from BALB/c mice infected with a bacterial pathogen. The InMAD serum, which is free of whole bacterial cells, is used to immunize syngeneic BALB/c mice. The resulting "InMAD immune serum" contains antibodies specific for the soluble microbial antigens present in sera from the infected mice. The InMAD immune serum is then used to probe blots of bacterial lysates or bacterial proteome arrays. Bacterial antigens that are reactive with the InMAD immune serum are precisely the antigens to target in an antigen immunoassay. By employing InMAD, we identified multiple circulating antigens that are secreted or shed during infection using Burkholderia pseudomallei and Francisella tularensis as model organisms. Potential diagnostic targets identified by the InMAD approach included bacterial proteins, capsular polysaccharide, and lipopolysaccharide. The InMAD technique makes no assumptions other than immunogenicity and has the potential to be a broad discovery platform to identify diagnostic targets from microbial pathogens. IMPORTANCE Effective treatment of microbial infection is critically dependent on early diagnosis and identification of the etiological agent. One means for rapid diagnosis is immunoassay for antigens that are shed into body fluids during infection. Immunoassays can be inexpensive, rapid, and adaptable to a point-of-care format. A major impediment to immunoassay for diagnosis of infectious disease is identification of appropriate antigen targets. This report describes a strategy that can be used for identification of microbial antigens that are shed into serum during infection by the biothreats Burkholderia pseudomallei and Francisella tularensis. Termed InMAD (in vivo microbial antigen discovery), the strategy has the potential for application to a broad spectrum of microbial pathogens. © 2011 Nuti et al.
ISSN: 21507511
Appears in Collections:Scopus 2011-2015

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