Implementación de la técnica de PCR enla identificación de Babesia ssp en equinos

Abstract

Babesia equi and Babesia caballi are intraerythrocytic protozoan parasites transmitted by ticks, causing equine babesiosis. Although the reference techniques recommended by USDA and OIE are IFAT and CF, they yields falsenegative results and dont let differentiate between babesias species. The implementation of polymerase chain reaction (PCR) as a direct technique for the identification and characterization of these parasites, without doubt enrich the clinical diagnosis of babesiosis. . For all those reasons in the present report was estandarizated PCR for the identification of B. equi and B. caballi. The proceeding included blood DNA extraction protocol and the PCR optimization for the reaction mix and the termocyclig program, with four primers asigned P1 and P2 for B. equi and P3 and P4 for B. caballi. Both amplified in a selective way a conserved regions from the 16S rRNA genes (rDNA) of 659 bp for B. equi and 664 bp for B. caballi. The minimal amount of DNA detected from positives controls was 0,1 ng/µl for B. equi and 1ng/µl for B. caballi. The primer set chosen do not produced amplifications fragments using others DNAs, like Toxoplasma gondii, Trypanosoma cruzi, Echinococcus granulosus, Fasciola hepatica. 77 horse blood specimens from Metropolitan Region, were tested for B. equi and B. caballi by PCR, 62 samples were negatives and 15 positives (14 to B. equi and 1 to B. caballi). With the aim of developing epidemiological studies, comparison PCR and microscopy, we are testing horse blood samples with suspicious of babesiosis.