title: Improved molecular technique for the differentiation of neotropical anopheline species.
abstract: We evaluated a PCR-RFLP of the ribosomal internal transcribed spacer 2 region (ITS2) to distinguish species of Anopheles commonly reported in the Amazon and validated this method using reared F1 offspring. The following species of Anopheles were used for molecular analysis: An. (Nys.) benarrochi, An. (Nys.) darlingi, An. (Nys.) nuneztovari, An. (Nys.) konderi, An. (Nys.) rangeli, and An. (Nys.) triannulatus sensu lato (s.l.). In addition, three species of the subgenus Anopheles, An. (Ano.) forattini, An. (Ano.) mattogrossensis, and An. (Ano.) peryassui were included for testing. Each of the nine species tested yielded diagnostic banding patterns. The PCR-RFLP method was successful in identifying all life stages including exuviae with small fractions of the sample. The assay is rapid and can be applied as an unbiased confirmatory method for identification of morphologic variants, disputed samples, imperfectly preserved specimens, and life stages from which taxonomic keys do not allow for definitive species determination.
authors.affiliation: University of California San Diego, La Jolla, California 92093, USA. ryanmatson@gmail.com
authors.forename: Ryan
authors.initials: R
authors.lastname: Matson
authors.forename: Carlos Tong
authors.initials: CT
authors.lastname: Rios
authors.forename: Cesar Banda
authors.initials: CB
authors.lastname: Chavez
authors.forename: Robert H
authors.initials: RH
authors.lastname: Gilman
authors.forename: David
authors.initials: D
authors.lastname: Florin
authors.forename: Victor Lopez
authors.initials: VL
authors.lastname: Sifuentes
authors.forename: Roldan Cardenas
authors.initials: RC
authors.lastname: Greffa
authors.forename: Pablo Peñataro
authors.initials: PP
authors.lastname: Yori
authors.forename: Roberto
authors.initials: R
authors.lastname: Fernandez
authors.forename: Daniel Velasquez
authors.initials: DV
authors.lastname: Portocarrero
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Vinetz
authors.forename: Margaret
authors.initials: M
authors.lastname: Kosek
mesh_terms: D000818:Animals; D000852:Anopheles / Q000145:classification* / Q000235:genetics*; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D013045:Species Specificity; D014329:Tropical Climate
chemical_list: D021903:DNA, Ribosomal Spacer