Expression and purification of Leptospira interrogans antigens. Application in serodiagnosis

VIARENGO, GASTÓN; VANASCO, NORMA BIBIANA; LOTTERSBERGER, JAVIER; GUERRERO, SERGIO ADRIÁN

Carlos Paz, Córdoba, Argentina

Congreso; XLIV Reunión Anual - Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB); 2008

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB)

Leptospirosis is a worldwide re-emerging zoonotic disease. The early and accurate detection of this infection is relevant in sanitary management. LipL32 is one of the most antigenic outer membrane proteins from pathogenic leptospiras. Another immunogen, Lp29, was reported as reactive since early stages of the illness. The aim of this work was the design of new antigens to develop ELISA tests for the detection of specific anti-leptospiral antibodies. We have cloned, expressed and purified the antigenic fraction of LipL32 protein and a quimeric protein consisting in the L29 antigen fused to the previously mentioned protein. These two recombinant proteins were evaluated by dot-blot and Western blot assays. Both assays were carried out by using specific hyperimmune rabbit sera and microaglutination (MAT) positive-negative human serum samples. LipL32 and the quimeric protein showed a good capacity to separate positive and negative samples by both immunoblot tests. The preliminary ELISA test results evidenced positive-negative differentiation capacity too. These results show a potential application of this recombinant designed proteins in the development of ELISA tests for detection of acute leptospirosis. Granted by UNL, CAI+D 2006; ANPCyT, PICTO´04 15-22427.