Introducing the ESAT-6 free IGRA, a companion diagnostic for TB vaccines based on ESAT-6

Morten Ruhwald, Lena de Thurah, Davis Kuchaka, Mostafa Rafaat Zaher, Ahmed M. Salman, Abdel-Rahman Abdel-Ghaffar, Faten Aly Shoukry, Sascha Wilk Michelsen, Bolette Soborg, Thomas Blauenfeldt, Stellah Mpagama, Søren T. Hoff, Else Marie Agger, Ida Rosenkrands, Claus Aagard, Gibson Kibiki, Nabila El-Sheikh, and Peter Andersen
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There is a need for an improved vaccine for tuberculosis. ESAT-6 is a cardinal vaccine antigen with unique properties and is included in several vaccine candidates in development. ESAT-6 is also the core antigen in the IFN-γ release assays (IGRA) used to diagnose latent infection, rendering IGRA tests unspecific after vaccination. This challenge has prompted the development of a companion diagnostic for ESAT-6 based vaccines, an ESAT-6 free IGRA. We screened a panel of seven potential new diagnostic antigens not recognized in BCG vaccinated individuals. Three highly recognized antigens EspC, EspF and Rv2348c were identified and combined with CFP10 in an ESAT-6 free antigen cocktail. The cocktail was prepared in a field-friendly format, lyophilized with heparin in ready-to-use vacutainer tubes. The diagnostic performance of the ESAT-6 free IGRA was determined in a cross-validation study. Compared IGRA, the ESAT-6 free IGRA induced a comparable magnitude of IFN-γ release, and the diagnostic performance was on par with Quantiferon (sensitivity 84% vs 79%; specificity 99% vs 97%). The comparable performance of the ESAT-6 free IGRA to IGRA suggests potential as companion diagnostic for ESAT-6 containing vaccines and as adjunct test for latent infection.