Detection and identification of Leishmania species within naturally infected sand flies in the Andean areas of Ecuador by a polymerase chain reaction

KATO, HIROTOMO; UEZATO, HIROSHI; KATAKURA, KEN; CALVOPINA, MANUEL; MARCO, JORGE DIEGO; BARROSO, PAOLA ANDREA; GOMEZ, EDUARDO; MIMORI, TATSUYUKI; KORENAGA, MASATAKA; IWATA, HIROYUKI; NONAKA, SHIGEO; HASHIGUCHI, YOSHIHISA

AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE

American Society of Tropical Medicine and Hygiene

Lugar: Cleveland, United States of America; Año: 2005 vol. 72 p. 87 - 93

0002-9637

Abstract. The surveillance of prevalent Leishmania and sand fly species in endemic areas is important for prediction of the risk and expansion of leishmaniasis. In this study, we developed a polymerase chain reaction (PCR)−based method for detection of Leishmania minicircle DNA within individual sand flies. Using this method, we detected minicircle DNAin 6 (3.3%) of 183 sand flies, while 5 (3.5%) of 143 were positive for Leishmania promastigotes in the same areas by microscopic examination. The species were identified as Leishmania (Leishmania) mexicana by nucleotide sequencing of the cytochrome b gene. Additionally, all the Leishmania-positive sand flies were identified as Lutzomyia ayacuchensisby the restriction enzyme digestion of the PCR-amplified 18S ribosomal RNA gene fragments. Since this combined method is relatively easy and can process a large number of samples, it will be a powerful tool for the rapid identification of prevalent sand fly and Leishmania species as well as monitoring the infection rate in sand fly populations in endemicareas.