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dc.contributor.authorPeter D. Burbeloen_US
dc.contributor.authorJason Kelleren_US
dc.contributor.authorJason Wagneren_US
dc.contributor.authorJames S. Klimaviczen_US
dc.contributor.authorAhmad Bayaten_US
dc.contributor.authorCraig S. Rhodesen_US
dc.contributor.authorBassirou Diarraen_US
dc.contributor.authorPloenchan Chetchotisakden_US
dc.contributor.authorYupin Suputtamongkolen_US
dc.contributor.authorSasisopin Kiertiburanakulen_US
dc.contributor.authorSteven M. Hollanden_US
dc.contributor.authorSarah K. Browneen_US
dc.contributor.authorSophia Siddiquien_US
dc.contributor.authorJoseph A. Kovacsen_US
dc.contributor.otherNational Institute of Dental and Craniofacial Researchen_US
dc.contributor.otherUniversity of Sciencesen_US
dc.contributor.otherKhon Kaen Universityen_US
dc.contributor.otherMahidol Universityen_US
dc.contributor.otherNational Institute of Allergy and Infectious Diseasesen_US
dc.contributor.otherNIH Clinical Centeren_US
dc.identifier.citationBMC Microbiology. Vol.15, No.1 (2015)en_US
dc.description.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.en_US
dc.rightsMahidol Universityen_US
dc.subjectImmunology and Microbiologyen_US
dc.titleSerological diagnosis of pulmonary Mycobacterium tuberculosis infection by LIPS using a multiple antigen mixtureen_US
Appears in Collections:Scopus 2011-2015

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