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Application of kDNA as a molecular marker to analyse Leishmania infantum diversity in Portugal
Authors:Cortes Sofia  Mauricio Isabel  Almeida Ana  Cristovão José Manuel  Pratlong Francine  Dedet Jean Pierre  Campino Lenea
Affiliation:

aUnidade de Leishmanioses, Centro Malária Outras Doenças Tropicais, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Rua da Junqueira, 96, 1349-008 Lisboa, Portugal

bPathogen Molecular Biology Unit, Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT, UK

cCentre National de Référence des Leishmania, Université de Montpellier, 163, rue Auguste Broussonet, 34090 Montpellier, France

Abstract:Around the Mediterranean basin Leishmania infantum is an important parasite causing canine leishmaniasis and visceral and cutaneous clinical forms in both immunocompetent and immunocompromised humans. Efficient monitoring and evaluation of epidemiology with discriminatory molecular markers are required. We investigated the genetic diversity of L. infantum in Portugal by polymerase chain amplification and restriction fragment length polymorphism analysis of kinetoplastid DNA, as molecular marker. We analysed 120 Portuguese isolates of L. infantum plus 16 other non-Portuguese isolates (as a reference group) from humans, dogs and sand flies. The Portuguese population showed a high degree of polymorphism with a total of 13 profiles identified. The predominant profile was A, which was only detected in the Portuguese samples.

The kinetoplastid DNA PCR-RFLP assay described here was suitable for use directly with biological samples and the profiles obtained were stable during long-term growth in vitro and in laboratory animals.

Keywords:Leishmania infantum   Epidemiology   Portugal   PCR-RFLP
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