Description

Background: Nontuberculous mycobacteria (NTM)-mediated infections are a growing cause of worldwide morbidity, but lack of rapid diagnostics for specific NTM species can delay the initiation of appropriate treatment regimens. We thus examined whether mass spectrometry analysis of an abundantly secreted mycobacterial

Background: Nontuberculous mycobacteria (NTM)-mediated infections are a growing cause of worldwide morbidity, but lack of rapid diagnostics for specific NTM species can delay the initiation of appropriate treatment regimens. We thus examined whether mass spectrometry analysis of an abundantly secreted mycobacterial antigen could identify specific NTM species.

Methods: We analyzed predicted tryptic peptides of the major mycobacterial antigen Ag85B for their capacity to distinguish Mycobacterium tuberculosis and three NTM species responsible for the majority of pulmonary infections caused by slow-growing mycobacterial species. Next, we analyzed trypsin-digested culture supernatants of these four mycobacterial species by liquid chromatography–tandem mass spectrometry (LC–MS/MS) to detect candidate species-specific Ag85B peptides, the identity of which were validated by LC–MS/MS performed in parallel reaction monitoring mode.

Results: Theoretical tryptic digests of the Ag85B proteins of four common mycobacterial species produced peptides with distinct sequences, including two peptides that could each identify the species origin of each Ag85B protein. LC–MS/MS analysis of trypsinized culture supernatants of these four species detected one of these species-specific signature peptides in each sample. Subsequent LC–MS/MS analyses confirmed these results by targeting these species-specific Ag85B peptides.

Conclusions: LC–MS/MS analysis of Ag85B peptides from trypsin-digested mycobacterial culture supernatants can rapidly detect and identify common mycobacteria responsible for most pulmonary infections caused by slow-growing mycobacteria, and has the potential to rapidly diagnose pulmonary infections caused by these mycobacteria through direct analysis of clinical specimens.

Reuse Permissions
  • Downloads
    PDF (1.1 MB)

    Details

    Title
    • Antigen 85B Peptidomic Analysis Allows Species-Specific Mycobacterial Identification
    Contributors
    Date Created
    2018-01-08
    Resource Type
  • Text
  • Collections this item is in
    Identifier
    • Digital object identifier: 10.1186/s12014-017-9177-6
    • Identifier Type
      International standard serial number
      Identifier Value
      1542-6416
    • Identifier Type
      International standard serial number
      Identifier Value
      1559-0275
    Note
    • The electronic version of this article is the complete one and can be found online at: https://clinicalproteomicsjournal.biomedcentral.com/articles/10.1186/s12014-017-9177-6

    Citation and reuse

    Cite this item

    This is a suggested citation. Consult the appropriate style guide for specific citation guidelines.

    Zhang, W., Shu, Q., Zhao, Z., Fan, J., Lyon, C. J., Zelazny, A. M., & Hu, Y. (2018). Antigen 85B peptidomic analysis allows species-specific mycobacterial identification. Clinical Proteomics, 15(1). doi:10.1186/s12014-017-9177-6

    Machine-readable links