Preliminary evaluation of a dual antigen ELISA to distinguish vaccinated from Leptospira infected horses.

Immunogenic proteins of Leptospira interrogans serovar Pomona type kennewicki (Lk) including Sph1, LigA, Hsp15 and LipL45 (Qlp42) are up-regulated in infected horses but are undetectable or expressed in trace amounts on cultured organisms. In contrast, LipL32 is abundant on cultured Lk and elicits infection antibody responses. The aim of this study was to develop an ELISA based on LipL32 or Lk sonicate and host-induced proteins to differentiate vaccine from infection serum antibody. IgG specific for recombinant Sph1, LigA, Lk90 (LigA; 379-1225 a.a), Hsp15, LipL45 and LipL32 of Lk were assayed in sera of horses infected naturally with Lk and before and after immunisation with serovar Pomona bacterin. Infection but not vaccine sera reacted strongly with Sph1, LigA and Lk90. LipL45 and Hsp15 reacted moderately with infection sera and weakly with vaccine sera. Lk sonicate and LipL32 reacted strongly with both infection and vaccine sera. As expected, culture-based vaccine failed to stimulate antibody to host-induced proteins. Therefore a dual antigen ELISA based on Lk sonicate or LipL32 combined with host-induced Sph1 and Lk90 will be valuable in differentiating infection from vaccine responses.