Identification of causative Leishmania species in Giemsa-stained smears prepared from patients with cutaneous leishmaniasis in Peru using PCR-RFLP

Acta Trop. 2016 Jun:158:83-87. doi: 10.1016/j.actatropica.2016.02.024. Epub 2016 Mar 2.

Abstract

A PCR-Restriction Fragment Length Polymorphism (RFLP) targeting the mannose phosphate isomerase gene was established to differentiate Leishmania species distributed near the Department of Huanuco, Peru. The technique was applied to 267 DNA samples extracted from Giemsa-stained smears of cutaneous lesions taken from patients suspected for cutaneous leishmaniasis in the area, and the present status of causative Leishmania species was identified. Of 114 PCR-amplified samples, 22, 19, 24 and 49 samples were identified to be infected by Leishmania (Viannia) braziliensis, L. (V.) peruviana, L. (V.) guyanensis, and a hybrid of L. (V.) braziliensis/L. (V.) peruviana, respectively, and the validity of PCR-RFLP was confirmed by sequence analysis. Since PCR-RFLP is simple and rapid, the technique will be a useful tool for the epidemiological study of leishmaniasis.

Keywords: Leishmania; Mannose phosphate isomerase; PCR-RFLP; Peru.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Azure Stains
  • Humans
  • Leishmania / classification*
  • Leishmania / genetics*
  • Leishmaniasis, Cutaneous / epidemiology*
  • Leishmaniasis, Cutaneous / genetics*
  • Peru / epidemiology
  • Polymerase Chain Reaction
  • Polymorphism, Restriction Fragment Length

Substances

  • Azure Stains