Molecular typing of sand fly species (Diptera, Psychodidae, Phlebotominae) from areas endemic for Leishmaniasis in Ecuador by PCR-RFLP of 18S ribosomal RNA gene.
Surveillance of the distribution of sand fly species is important for prediction of the risk and expansion of Leishmania infection in endemic and surrounding areas. In the present study, a simple and reliable method of typing New World Lutzomyia species circulating in endemic areas in Ecuador was established by using polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) technique. PCR-RFLP of 18S ribosomal RNA (rRNA) genes with the restriction enzyme AfaI and subsequently HinfI successfully identified seven sand fly species in nine endemic areas in Ecuador. Although intraspecific genetic-diversity affecting the RFLP-patterns was detected in a species, the patterns were species specific. The method promises to be a powerful tool for the classification of New World Lutzomyia species.
Department of Veterinary Hygiene, Faculty of Agriculture, Yamaguchi University, Yamaguchi, Japan.
Yoshimi
Y
Terayama
Hirotomo
H
Kato
Eduardo A
EA
Gomez
Hiroshi
H
Uezato
Manuel
M
Calvopiña
Hiroyuki
H
Iwata
Yoshihisa
Y
Hashiguchi
D000818:Animals; D016000:Cluster Analysis; D017931:DNA Primers / Q000235:genetics; D004484:Ecuador / Q000453:epidemiology; D014644:Genetic Variation; D007896:Leishmaniasis / Q000453:epidemiology*; D010802:Phylogeny; D012150:Polymorphism, Restriction Fragment Length / Q000235:genetics; D011576:Psychodidae / Q000145:classification / Q000235:genetics*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D013045:Species Specificity
D017931:DNA Primers; D012337:RNA, Ribosomal, 18S