Improved Molecular Technique for the Differentiation of Neotropical Anopheline Species

Ryan Matson University of California San Diego, La Jolla, California; Navy Medical Research Center Detachment, Iquitos, Peru; A.B. Prisma, Iquitos, Peru; Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Navy Medical Research Center Detachment, Lima, Peru; Ministry of Health, Loreto, Peru

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Carlos Tong Rios University of California San Diego, La Jolla, California; Navy Medical Research Center Detachment, Iquitos, Peru; A.B. Prisma, Iquitos, Peru; Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Navy Medical Research Center Detachment, Lima, Peru; Ministry of Health, Loreto, Peru

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Cesar Banda Chavez University of California San Diego, La Jolla, California; Navy Medical Research Center Detachment, Iquitos, Peru; A.B. Prisma, Iquitos, Peru; Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Navy Medical Research Center Detachment, Lima, Peru; Ministry of Health, Loreto, Peru

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Robert H. Gilman University of California San Diego, La Jolla, California; Navy Medical Research Center Detachment, Iquitos, Peru; A.B. Prisma, Iquitos, Peru; Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Navy Medical Research Center Detachment, Lima, Peru; Ministry of Health, Loreto, Peru

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David Florin University of California San Diego, La Jolla, California; Navy Medical Research Center Detachment, Iquitos, Peru; A.B. Prisma, Iquitos, Peru; Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Navy Medical Research Center Detachment, Lima, Peru; Ministry of Health, Loreto, Peru

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Victor Lopez Sifuentes University of California San Diego, La Jolla, California; Navy Medical Research Center Detachment, Iquitos, Peru; A.B. Prisma, Iquitos, Peru; Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Navy Medical Research Center Detachment, Lima, Peru; Ministry of Health, Loreto, Peru

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Roldan Cardenas Greffa University of California San Diego, La Jolla, California; Navy Medical Research Center Detachment, Iquitos, Peru; A.B. Prisma, Iquitos, Peru; Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Navy Medical Research Center Detachment, Lima, Peru; Ministry of Health, Loreto, Peru

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Pablo Peñataro Yori University of California San Diego, La Jolla, California; Navy Medical Research Center Detachment, Iquitos, Peru; A.B. Prisma, Iquitos, Peru; Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Navy Medical Research Center Detachment, Lima, Peru; Ministry of Health, Loreto, Peru

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Roberto Fernandez University of California San Diego, La Jolla, California; Navy Medical Research Center Detachment, Iquitos, Peru; A.B. Prisma, Iquitos, Peru; Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Navy Medical Research Center Detachment, Lima, Peru; Ministry of Health, Loreto, Peru

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Daniel Velasquez Portocarrero University of California San Diego, La Jolla, California; Navy Medical Research Center Detachment, Iquitos, Peru; A.B. Prisma, Iquitos, Peru; Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Navy Medical Research Center Detachment, Lima, Peru; Ministry of Health, Loreto, Peru

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Joseph M. Vinetz University of California San Diego, La Jolla, California; Navy Medical Research Center Detachment, Iquitos, Peru; A.B. Prisma, Iquitos, Peru; Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Navy Medical Research Center Detachment, Lima, Peru; Ministry of Health, Loreto, Peru

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Margaret Kosek University of California San Diego, La Jolla, California; Navy Medical Research Center Detachment, Iquitos, Peru; A.B. Prisma, Iquitos, Peru; Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland; Navy Medical Research Center Detachment, Lima, Peru; Ministry of Health, Loreto, Peru

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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.

Author Notes

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