Evaluation of two cocktails containing ESAT-6, CFP-10 and Rv-3615c in the intradermal test and the interferon-γ assay for diagnosis of bovine tuberculosis

Prev Vet Med. 2012 Jun 1;105(1-2):149-54. doi: 10.1016/j.prevetmed.2012.02.007. Epub 2012 Mar 3.


The intradermal tuberculin tests and the interferon-gamma (IFN-γ) assay are the principal tests used worldwide for the ante-mortem diagnosis of bovine tuberculosis. The conventional reagent currently in use in these tests is purified protein derivative (PPD) tuberculin obtained from Mycobacterium bovis culture. The components of PPD are poorly characterized and difficult to standardize. To overcome this issue, antigens specific to the Mycobacterium tuberculosis complex are being studied. Here we have assessed the biological potency of ESAT-6, CFP-10 and Rv-3615c presented as peptide or recombinant protein cocktails in comparison with the standard bovine PPD used routinely in Spanish eradication campaigns. The study was performed in cattle (n=23) from a herd with natural M. bovis infection. Animals were simultaneously injected with PPD and the peptide and protein cocktails. The percentages of cattle reacting positively to single intradermal test were 60.9% (bovine PPD), 47.8% (peptide cocktail) and 60.9% (protein cocktail), with no significant difference between the actual skin fold thickness increases (p>0.05). The IFN-γ assay detected 60.9% of animals when stimulation was performed with bovine PPD, but decreased to 52.2% when stimulation was performed with the peptide cocktail and to 47.8% when stimulation was performed with the protein cocktail. However, no significant differences were found between IFN-γ responder frequencies (p>0.05). These results show a potential use of these defined reagents for in vivo tuberculosis diagnosis.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Bacterial
  • Bacterial Proteins
  • Cattle
  • Interferon-gamma Release Tests / methods*
  • Interferon-gamma Release Tests / veterinary
  • Intradermal Tests / methods*
  • Intradermal Tests / veterinary
  • Mycobacterium bovis / immunology*
  • Recombinant Proteins*
  • Sensitivity and Specificity
  • Tuberculin Test / methods*
  • Tuberculin Test / veterinary
  • Tuberculosis, Bovine / diagnosis*


  • Antigens, Bacterial
  • Bacterial Proteins
  • Recombinant Proteins