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

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 DNA in 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 ayacuchensis by 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 endemic areas.Fil: Kato, Hirotomo. Yamaguchi University; JapónFil: Uezato, Hiroshi. University of the Ryukyus; JapónFil: Katakura, Ken. Hokkaido University; JapónFil: Calvopina, Manuel. Kochi University. Kochi Medical School; JapónFil: Marco, Jorge Diego. Kochi University. Kochi Medical School; Japón. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; ArgentinaFil: Barroso, Paola Andrea. Kochi University. Kochi Medical School; Japón. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Patología Experimental. Universidad Nacional de Salta. Facultad de Ciencias de la Salud. Instituto de Patología Experimental; ArgentinaFil: Gomez, Eduardo. Universidad Católica de Guayaquil; EcuadorFil: Mimori, Tatsuyuki. Kumamoto University; JapónFil: Korenaga, Masataka. Kochi University. Kochi Medical School; JapónFil: Iwata, Hiroyuki. Yamaguchi University; JapónFil: Nonaka, Shigeo. University ok the Ryukyus; JapónFil: Hashiguchi, Yoshihisa. Kochi University. Kochi Medical School; Japó