title: 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.
authors.affiliation: Department of Veterinary Hygiene, Faculty of Agriculture, Yamaguchi University, Yamaguchi, Japan. katoh@yamaguchi-u.ac.jp
authors.forename: Hirotomo
authors.initials: H
authors.lastname: Kato
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Uezato
authors.forename: Ken
authors.initials: K
authors.lastname: Katakura
authors.forename: Manuel
authors.initials: M
authors.lastname: Calvopiña
authors.forename: Jorge D
authors.initials: JD
authors.lastname: Marco
authors.forename: Paola A
authors.initials: PA
authors.lastname: Barroso
authors.forename: Eduardo A
authors.initials: EA
authors.lastname: Gomez
authors.forename: Tatsuyuki
authors.initials: T
authors.lastname: Mimori
authors.forename: Masataka
authors.initials: M
authors.lastname: Korenaga
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Iwata
authors.forename: Shigeo
authors.initials: S
authors.lastname: Nonaka
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Hashiguchi
mesh_terms: D000818:Animals; D018105:DNA, Kinetoplast / Q000032:analysis* / Q000235:genetics; D016054:DNA, Protozoan / Q000032:analysis* / Q000235:genetics; D004484:Ecuador; D007303:Insect Vectors / Q000469:parasitology; D007891:Leishmania / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D007896:Leishmaniasis / Q000635:transmission; D016133:Polymerase Chain Reaction / Q000379:methods; D011576:Psychodidae / Q000469:parasitology*; D012337:RNA, Ribosomal, 18S / Q000032:analysis*
chemical_list: D018105:DNA, Kinetoplast; D016054:DNA, Protozoan; D012337:RNA, Ribosomal, 18S