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Title: Serological diagnosis of pulmonary Mycobacterium tuberculosis infection by LIPS using a multiple antigen mixture
Authors: Peter D. Burbelo
Jason Keller
Jason Wagner
James S. Klimavicz
Ahmad Bayat
Craig S. Rhodes
Bassirou Diarra
Ploenchan Chetchotisakd
Yupin Suputtamongkol
Sasisopin Kiertiburanakul
Steven M. Holland
Sarah K. Browne
Sophia Siddiqui
Joseph A. Kovacs
National Institute of Dental and Craniofacial Research
University of Sciences
Khon Kaen University
Mahidol University
National Institute of Allergy and Infectious Diseases
NIH Clinical Center
Keywords: Immunology and Microbiology;Medicine
Issue Date: 8-Oct-2015
Citation: BMC Microbiology. Vol.15, No.1 (2015)
Abstract: © 2015 Burbelo et al. Background: There is an urgent need for a simple and accurate test for the diagnosis of human Mycobacterium tuberculosis, the infectious agent causing tuberculosis (TB). Here we describe a serological test based on light emitting recombinant proteins for the diagnosis of pulmonary Mycobacterium tuberculosis infection. Methods: Luciferase Immunoprecipitation Systems (LIPS), a fluid-phase immunoassay, was used to examine antibody responses against a panel of 24 different M. tuberculosis proteins. Three different strategies were used for generating the constructs expressing the recombinant fusion M. tuberculosis proteins with luciferase: synthetic gene synthesis, Gateway recombination cloning, and custom PCR synthesis. A pilot cohort of African pulmonary TB patients was used for initial antibody screening and confirmatory studies with selected antigens were performed with a cohort from Thailand and healthy US blood donors. In addition to testing M. tuberculosis antigens separately, a mixture that tested seven antigens simultaneously was evaluated for diagnostic performance. Results: LIPS testing of a pilot set of serum samples from African pulmonary TB patients identified a potential subset of diagnostically useful M. tuberculosis antigens. Evaluation of a second independent cohort from Thailand validated highly significant antibody responses against seven antigens (PstS1, Rv0831c, FbpA, EspB, bfrB, HspX and ssb), which often showed robust antibody levels up to 50- to 1000-fold higher than local community controls. Marked heterogeneity of antibody responses was observed in the patients and the combined results demonstrated 73.5 % sensitivity and 100 % specificity for detection of pulmonary TB. A LIPS test simultaneously employing the seven M. tuberculosis antigen as a mixture matched the combined diagnostic performance of the separate tests, but showed an even higher diagnostic sensitivity (90 %) when a cut-off based on healthy US blood donors was used. Conclusion: A LIPS immunoassay employing multiple M. tuberculosis antigens shows promise for the rapid and quantitative serological detection of pulmonary TB.
ISSN: 14712180
Appears in Collections:Scopus 2011-2015

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