Resumen

GONZALEZ M, Eglys B et al. Standarization of a polimerase chain reaction technique for the diagnosis of animal trypanosomiasis caused by  Trypanosoma evansi. Agronomía Trop. [online]. 2006, vol.56, n.4, pp.495-500. ISSN 0002-192X.

There are several methods to diagnose this illness, equine trypanosomiasis,caused by Trypanosoma evansi and their sensibility varies depending on thedisease stage. Recently, molecular methods, like PCR, based on DNA detectionhave improved the diagnosis of this disease. The purpose of this work was tostandardize, differentiate and evaluate the PCR technique for the diagnosis of T. evansi in an experimental murine model. Mice were infected with TEVA1.After 48 hours they were evaluated using the parasitological methods of Brener’squantification and the Woo haematocrit centrifuge technique. For moleculardiagnosis, genomic DNA was purified and amplified by PCR using primersspecific for T. evansi: ESAG 6/7, TEV1/2 y TBR1/2. All primers rendered theexpected product of amplification, except when non infected animals were used.Primers ESAG 6/7 showed a sensibility of 1ng using DNA from purifiedparasites and of 10ng using DNA from whole blood samples. From the total ofmice infected, 47% (n=8) were positive by Brener’s method, 70% (n=12) byWoo’s method, and all (100%) by the PCR technique. This result indicates thatthe PCR technique is much more efficient and sensitive than the parasitologicalmethods used for diagnosing T. evansi in a murine model when there is low concentration of parasites.

Palabras clave : Trypanosoma evansi; diagnosis; PCR.