Amplified Fragment Length Polymorphisms Reveals High Intraspecific Variability in Field Isolates of Leishmania panamensis

Order of Publishing in Issue: 
12
Volume :5
Issue :2
April, 2011
Page No: 
1183-1192
Authors: 
Carlos M. Restrepo[1], Efraín Pérez Lao [1], Carolina De La Guardia [1], Octavio E. Sousa [2], José E. Calzada [3] and Ricardo Lleonart [1]*
Address: 
[1] Instituto de Investigaciones Científicas y Servicios de Alta Tecnología (INDICASAT-AIP), Panamá
Address: 
[2] Centro de Investigaciones y Diagnóstico de Enfermedades Parasitarias (CIDEP), Facultad de Medicina, Universidad de Panamá, Panamá
Address: 
[3] Instituto Conmemorativo GORGAS de Estudios de la Salud, Panamá

Abstract : Leishmania parasites cause leishmaniasis, a potentially deadly re-emergent disease that affects millions throughout the world. In Panama,the disease is showing an increasing trend, with estimates of thousands of new cases every year.  The main manifestations are the cutaneous andmucocutaneous forms. Genetic variability studies in Leishmania are extremely important to define key elements of the eco-epidemiology of thedisease. However, few studies have addressed this issue in Panama, and these have been based mainly on kinetoplastid DNA RFLP. The amplifiedfragment length polymorphisms (AFLP) is a very efficient technique for rapid detection of genetic variability, particularly useful on organisms withoutsequenced genomes. Although this technique has been used successfully on many species, including several protozoa, its use for studying geneticvariability in Leishmania parasites is just in its beginnings. We have optimized and used AFLP to address genetic diversity in Leishmaniapanamensis, a poorly studied member of the Viannia subgenus. We have found that this technique is able to generate high numbers ofpeaks when low selective EcoRI and MseI primers were used (+0, +1, +2 series). Additionally, we have found that an importantproportion of those alleles, up to 57% for some primer combinations, are polymorphic. Some of these alleles are potentially useful to rapidlydistinguish L. panamensis and L. guyanensis, the two most genetically similar species of the subgenus. The AFLP was an efficient techniqueto screen the Leishmania panamensis genome for polymorphisms, allowing the rapid detection of hundreds of polymorphic alleles.

Keywords: 
AFLP, Leishmania, Genetic Variability, Polymorphisms
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