Use of AFLP to study the genetic variability of Leishmania sp

Abstract

AFLP is a very efficient technique for rapid detection of genetic variability, particularly useful on organisms without sequenced genomes. The objective of this study was to assess the amplified fragment length polymorphisms (AFLP) for testing genetic diversity in Leishmania sp. Leishmania species used for this study were representative of the Viannia and Leishmania subgenus. Fluorescent AFLP kits (Applied Biosystems, USA) were used for the whole procedure. Detection and sizing of peaks were conducted with GeneMarker v. 1.91 software (Softgenetics, USA). Then error rate was estimated for every possible pair of replicates using ten replicates from a single L. panamensis promastigotes culture. We have found that this technique is able to generate high numbers of peaks when low selective EcoRI and MseI primers were used (+0, +1, +2 series). Additionally, we have found that up to 86,9% of the primer combinations are polymorphic. Several fragments were apparently specific for L panamensis, L. (Viannia) and L. (Leishmania). The error rates estimated from every possible pair of replicates ranged from 0% to 6%. Cluster analysis results are in agreement with the current known taxonomy of Leishmania species. In conclusion, AFLP is a molecular fingerprinting technique showing promising potential to explore genetic diversity at Leishmania sp., both at intraspecific and interspecific levels. Its application will allow researchers to look deeper into new polymorphisms and possibly correlate them to ecoepidemiologically relevant traits and other important aspects of the biology of the parasite.