Serological diagnosis of pulmonary Mycobacterium tuberculosis infection by LIPS using a multiple antigen mixture

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

Research output: Contribution to journalArticlepeer-review

15 Scopus citations


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.

Original languageEnglish (US)
Article number205
JournalBMC microbiology
Issue number1
StatePublished - Oct 8 2015
Externally publishedYes


  • Antibodies
  • Latent tuberculosis infection (LTBI)
  • Luciferase immunoprecipitation systems (LIPS)
  • Mycobacterium tuberculosis
  • Pulmonary TB
  • Serology

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)


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