Improved molecular technique for the differentiation of neotropical anopheline species.
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.
Matson
Ryan
R
University of California San Diego, La Jolla, California 92093, USA. ryanmatson@gmail.com
Rios
Carlos Tong
CT
Chavez
Cesar Banda
CB
Gilman
Robert H
RH
Florin
David
D
Sifuentes
Victor Lopez
VL
Greffa
Roldan Cardenas
RC
Yori
Pablo Peñataro
PP
Fernandez
Roberto
R
Portocarrero
Daniel Velasquez
DV
Vinetz
Joseph M
JM
Kosek
Margaret
M
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
D021903:DNA, Ribosomal Spacer