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...

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Published inThe American journal of tropical medicine and hygiene Vol. 78; no. 3; pp. 492 - 498
Main Authors Matson, Ryan, Rios, Carlos Tong, Chavez, Cesar Banda, Gilman, Robert H, Florin, David, Sifuentes, Victor Lopez, Greffa, Roldan Cardenas, Yori, Pablo Penataro, Fernandez, Roberto, Portocarrero, Daniel Velasquez, Vinetz, Joseph M, Kosek, Margaret
Format Journal Article
LanguageEnglish
Published Lawrence, KS ASTMH 01.03.2008
Allen Press
Subjects
Animals
Anopheles
Anopheles - classification
Anopheles - genetics
Biological and medical sciences
DNA, Ribosomal Spacer - genetics
Infectious diseases
Medical sciences
Polymerase Chain Reaction - methods
Polymorphism, Restriction Fragment Length
Species Specificity
Tropical Climate
Insecta
Polymerase chain reaction
Anopheles
Restriction fragment length polymorphism
Arthropoda
Culicidae
Genetics
Tropical medicine
Technique
Invertebrata
Molecular biology
Differentiation
Diptera
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Summary: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.
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ISSN:0002-9637
1476-1645
DOI:10.4269/ajtmh.2008.78.492