Tuberculin skin testing boosts interferon gamma responses to DIVA reagents in Mycobacterium bovis-Infected cattle

Gareth J. Jones, Mick Coad, Bhagwati Khatri, Javier Bezos, Natalie A. Parlane, Bryce M. Buddle, Bernardo Villarreal-Ramos, R. Glyn Hewinson, H. Martin Vordermeier

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Mycobacterium bovis BCG vaccination sensitizes cattle to bovine tuberculin, which compromises the use of the current bovine tuberculosis (TB) surveillance tests. Although the performance of a blood test (that utilizes antigens expressed by Mycobacterium bovis but not by BCG) capable of discriminating infected from vaccinated animals (DIVA interferon gamma test [DIT]) has been evaluated in naturally infected TB field reactors, there is a need to perform similar analysis in a BCG-vaccinated M. bovis-infected population. Furthermore, we explored different scenarios under which a DIT may be implemented alongside BCG vaccination: (i) serial testing to resolve potential false-positive skin test results or (ii) a standalone test to replace the single intradermal comparative cervical tuberculin (SICCT) skin test. Our results demonstrated significantly better relative test sensitivity when the DIT was evaluated in a serial test scenario. Direct comparison of pre-and post-skin test blood samples revealed that the SICCT test induced significant boosting of the gamma interferon response in M. bovis-infected animals to both the ESAT-6-CFP-10 and Rv3615c peptide cocktails that comprise the DIT, which persisted for the ESAT-6-CFP-10 reagent for at least 14 days. Importantly, no similar boosting effects were observed in noninfected BCG vaccinates, suggesting that DIVA blood testing after a recent skin test would have minimal impact on test specificity.

Original languageEnglish
Article numbere00551
Number of pages9
JournalClinical and Vaccine Immunology
Issue number5
Publication statusPublished - 05 May 2017


  • Blood test
  • Bovine tuberculosis
  • DIVA
  • Gamma interferon
  • Mycobacterium bovis
  • Tuberculin skin test


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