Identification of Proteins from Tuberculin Purified Protein Derivative (PPD) with Potential for TB Diagnosis Using Bioinformatics Analysis

  • Sibele Borsuk
  • Fabiana Kommling Seixas
  • Daniela Fernandes Ramos
  • Caroline Rizzi
  • Odir Antonio Dellagostin
Part of the Lecture Notes in Computer Science book series (LNCS, volume 5676)


The PPD is widely used for diagnosis of bovine tuberculosis, however it lacks specificity and sensitivity, generally attributed the cross-reactions with antigens shared by other Mycobacterium species. These highlight the necessity of better diagnostic tools to detect tuberculosis in cattle. We have identified proteins present in PPD preparations (avium and bovine PPD) by LC-MS/MS (Liquid Chromatography/Mass Spectrometry/Mass Spectrometry). A total of 169 proteins were identified. From these, four PPD proteins identified in bovine PPD and absent in avium PPD (Mb1961c, Mb2898, Mb2900 and Mb2965c) were select for bioinformatics analysis to identify an antigen with potential for TB diagnosis. We identified Mb1961c, a secreted protein, which has low identity proteins found in environmental mycobacteria. This protein could be used for the development of a diagnostic test or a synthetic PPD for bovine tuberculosis.


bioinformatics analysis LC-MS/MS bovine tuberculosis diagnostic 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    WHO: Global Tuberculosis Control. WHO report. WHO/CDS/TB/2002 (2002)Google Scholar
  2. 2.
    Hewinson, R.G., Vordermeier, H.M., Buddle, B.M.: Use of the bovine model of tuberculosis for the development of improved vaccines and diagnostics. Tuberculosis (Edinb) 83(1-3), 119–130 (2003)CrossRefGoogle Scholar
  3. 3.
    Daniel, T.M., Janicki, B.W.: Mycobacterial antigens: a review of their isolation, chemistry, and immunological properties. Microbiol. Rev. 42(1), 84–113 (1978)PubMedPubMedCentralGoogle Scholar
  4. 4.
    Young, D.B.: Heat-shock proteins: immunity and autoimmunity. Curr. Opin. Immunol. 4(4), 396–400 (1992)CrossRefPubMedGoogle Scholar
  5. 5.
    Vordermeier, H.M., Chambers, M.A., Buddle, B.M., Pollock, J.M., Hewinson, R.G.: Progress in the development of vaccines and diagnostic reagents to control tuberculosis in cattle. Vet. J. 171(2), 229–244 (2006)CrossRefPubMedGoogle Scholar
  6. 6.
    Klausen, J., Magnusson, M., Andersen, A.B., Koch, C.: Characterization of purified protein derivative of tuberculin by use of monoclonal antibodies: isolation of a delayed-type hypersensitivity reactive component from M. tuberculosis culture filtrate. Scand. J. Immunol. 40(3), 345–349 (1994)CrossRefPubMedGoogle Scholar
  7. 7.
    Rowland, S.S., Ruckert, J.L., Cummings, P.J.: Low molecular mass protein patterns in mycobacterial culture filtrates and purified protein derivatives. FEMS Immunol. Med. Microbiol. 23(1), 21–25 (1999)CrossRefPubMedGoogle Scholar
  8. 8.
    ANON.: The development of improved tests for the diagnosis of Mycobacterium bovis infection in cattle. Report of the Independent Scientific Group Vaccine Scoping Sub-Committee. Defra Publications, London (2006)Google Scholar
  9. 9.
    Vordermeier, H.M., Cockle, P.C., Whelan, A., et al.: Development of diagnostic reagents to differentiate between Mycobacterium bovis BCG vaccination and M. bovis infection in cattle. Clin. Diagn. Lab Immunol. 6(5), 675–682 (1999)PubMedPubMedCentralGoogle Scholar
  10. 10.
    Vordermeier, H.M., Whelan, A., Cockle, P.J., Farrant, L., Palmer, N., Hewinson, R.G.: Use of synthetic peptides derived from the antigens ESAT-6 and CFP-10 for differential diagnosis of bovine tuberculosis in cattle. Clin. Diagn. Lab Immunol. 8(3), 571–578 (2001)PubMedPubMedCentralGoogle Scholar
  11. 11.
    Gluckman, S.J.: Mycobacterium marinum. Clin. Dermatol. 13(3), 273–276 (1995)CrossRefPubMedGoogle Scholar
  12. 12.
    Lyashchenko, K.P., Pollock, J.M., Colangeli, R., Gennaro, M.L.: Diversity of antigen recognition by serum antibodies in experimental bovine tuberculosis. Infect. Immun. 66(11), 5344–5349 (1998)PubMedPubMedCentralGoogle Scholar
  13. 13.
    Manca, C., Lyashchenko, K., Wiker, H.G., Usai, D., Colangeli, R., Gennaro, M.L.: Molecular cloning, purification, and serological characterization of MPT63, a novel antigen secreted by Mycobacterium tuberculosis. Infect. Immun. 65(1), 16–23 (1997)PubMedPubMedCentralGoogle Scholar
  14. 14.
    Buddle, B.M., Parlane, N.A., Keen, D.L., et al.: Differentiation between Mycobacterium bovis BCG-vaccinated and M. bovis-infected cattle by using recombinant mycobacterial antigens. Clin. Diagn. Lab Immunol. 6(1), 1–5 (1999)PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2009

Authors and Affiliations

  • Sibele Borsuk
    • 1
  • Fabiana Kommling Seixas
    • 1
  • Daniela Fernandes Ramos
    • 1
  • Caroline Rizzi
    • 1
  • Odir Antonio Dellagostin
    • 1
  1. 1.Centro de BiotecnologiaUniversidade Federal de PelotasPelotasBrazil

Personalised recommendations