Calmodulin Polymerase Chain Reaction-Restriction Fragment Length Polymorphism for Leishmania Identification and Typing

Am J Trop Med Hyg. 2016 Aug 3;95(2):383-7. doi: 10.4269/ajtmh.15-0709. Epub 2016 Jun 27.

Abstract

A precise identification of Leishmania species involved in human infections has epidemiological and clinical importance. Herein, we describe a preliminary validation of a restriction fragment length polymorphism assay, based on the calmodulin intergenic spacer region, as a tool for detecting and typing Leishmania species. After calmodulin amplification, the enzyme HaeIII yielded a clear distinction between reference strains of Leishmania mexicana, Leishmania amazonensis, Leishmania infantum, Leishmania lainsoni, and the rest of the Viannia reference species analyzed. The closely related Viannia species: Leishmania braziliensis, Leishmania panamensis, and Leishmania guyanensis, are separated in a subsequent digestion step with different restriction enzymes. We have developed a more accessible molecular protocol for Leishmania identification/typing based on the exploitation of part of the calmodulin gene. This methodology has the potential to become an additional tool for Leishmania species characterization and taxonomy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calmodulin / genetics*
  • DNA, Protozoan / genetics
  • Humans
  • Leishmania braziliensis / classification*
  • Leishmania braziliensis / genetics
  • Leishmania braziliensis / isolation & purification
  • Leishmania guyanensis / classification*
  • Leishmania guyanensis / genetics
  • Leishmania guyanensis / isolation & purification
  • Leishmania infantum / classification*
  • Leishmania infantum / genetics
  • Leishmania infantum / isolation & purification
  • Leishmania mexicana / classification*
  • Leishmania mexicana / genetics
  • Leishmania mexicana / isolation & purification
  • Leishmaniasis / diagnosis
  • Leishmaniasis / parasitology
  • Molecular Typing / methods*
  • Polymerase Chain Reaction / methods
  • Polymorphism, Restriction Fragment Length
  • Protozoan Proteins / genetics

Substances

  • Calmodulin
  • DNA, Protozoan
  • Protozoan Proteins