Since the 1980s, the main objective

of VL diagnostics development has been to replace the visual identification of parasites in tissue, a technique that is invasive and requires considerable expertise. Moreover, the use of whole parasite extracts in the serologic tests is limited because of the low reproducibility and low specificity values obtained. Several serological tests have been developed, but none of them are specific for VL, although they have proved to be useful in combination with a clinical case definition (20). So, to obtain a specific diagnosis for VL, some purified recombinant Leishmania antigens have been proposed (21). The present study describes the expression, isolation and purification of two recombinant proteins from L. chagasi, rLci2B and rLci1A that were previously selected from a cDNA library, followed by standardization of an ELISA using South America canine sera, to contribute selleck products to the diagnostic of dog’s infection provoked by the protozoa parasite. Both proteins were produced in Escherichia coli, and their sequences were registered by the National Institute of Industrial Property – Brazil (INPI) under paragraph PI0900961-2, with the title: ‘Use of antigens of Leishmania in methods for diagnosis, therapy and vaccine for leishmaniasis’. The rLci2B protein has homology with a parasite cytoskeleton protein kinesin, while rLci1A has homology with the heat shock protein 70 (HSP70). The

HSP70 family belongs to Rutecarpine a class of proteins highly conserved throughout evolution and has an immunogenic activity

(22). Antibodies to specific recombinant antigens such find more as heat shock protein 70 (rHSP70) and kinesin K39 (rK39) have also been shown to be good markers to detect infection (23). Thus, this study will be useful to expand the panel of recombinant antigens for the development of more sensitive and specific serodiagnostic tests for this disease. All reagents used were of analytical grade. Distilled water was filtered and deionized using a Millipore water purification system. The sera used in this study were collected from domestic dogs from three Brazilian regions: Northeast, Southeast and Midwest. The serum samples were collected by venipuncture by veterinaries of three research centres: Centro de Pesquisa Aggeu Magalhães, Pernambuco; Instituto de Pesquisa Evandro Chagas, Rio de Janeiro and Centro Gonçalo Muniz, Bahia, between 2006 and 2008. The sera were stored at −70°C and transferred to our laboratory. The panel of 56 negative sera used for determining the cut-off came from the state of Rio de Janeiro. The multicentre panel of 119 negative sera used in the assay came from the Southeast (79), Midwest (26) and Northeast (14) regions of Brazil. The multicentre panel of 138 positive sera used in the challenge originated in the Southeast (38), Midwest (46) and Northeast (54) regions of Brazil. The panel of 86 negative sera for L.