Identificación de los alelos del Nramp1 asociados a la resistencia a Leishmaniasis en caninos (Canis lupus familiaris) mediante ARMS-PCR

Abstract

Canine Leishmaniasis is a zoonotic disease of great public health importance, especially in endemic areas such as Ecuador. Previous studies have shown that the Nramp1 gene plays a crucial role in resistance and susceptibility to intracellular diseases through macrophage regulation. This study aimed to identify Nramp1 alleles associated with resistance in canines (Canis lupus familiaris) from urban and rural areas of Cuenca, using the ARMS-PCR technique. Eighty-five DNA samples were processed, designing specific primers for the A4549G (intron 6), C4859T (exon 8), and C8542T (intron 13) polymorphisms. Of these, only the A4549G allele was successfully amplified, revealing a genotypic distribution of 89.41% AA homozygotes (associated with susceptibility) and 10.59% GA heterozygotes (partial resistance); no GG homozygotes were detected. The results indicate that the G allele in A4549G could be linked to some degree of protection against Leishmania spp., although its low frequency (4.71%) suggests limited selective pressure within the studied population. The ARMS-PCR technique proved to be efficient for genotyping this SNP, although conditions had to be adjusted for other polymorphisms.