TY - JOUR
T1 - Improved molecular technique for the differentiation of neotropical anopheline species
AU - Matson, Ryan
AU - Rios, Carlos Tong
AU - Chavez, Cesar Banda
AU - Gilman, Robert H.
AU - Florin, David
AU - Sifuentes, Victor Lopez
AU - Greffa, Roldan Cardenas
AU - Yori, Pablo Peñataro
AU - Fernandez, Roberto
AU - Portocarrero, Daniel Velasquez
AU - Vinetz, Joseph M.
AU - Kosek, Margaret
PY - 2008/3
Y1 - 2008/3
N2 - 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.
AB - 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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U2 - 10.4269/ajtmh.2008.78.492
DO - 10.4269/ajtmh.2008.78.492
M3 - Article
C2 - 18337348
AN - SCOPUS:42949102012
SN - 0002-9637
VL - 78
SP - 492
EP - 498
JO - American Journal of Tropical Medicine and Hygiene
JF - American Journal of Tropical Medicine and Hygiene
IS - 3
ER -