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© Research
Publication : Journal of Medical Entomology

Discrimination of 15 Amazonian Anopheline Mosquito Species by Polymerase Chain Reaction—Restriction Fragment Length Polymorphism

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Journal of Medical Entomology - 01 May 2022

S. B. Vezenegho, J. Issaly, R. Carinci, P. Gaborit, Romain Girod, Isabelle Dusfour, S. Briolant

Link to HAL – amu-03678991

Link to DOI – 10.1093/jme/tjac008

Journal of Medical Entomology, 2022, 59 (3), pp.1060-1064. ⟨10.1093/jme/tjac008⟩

Precise identification of anopheline species is paramount for incrimination of malaria vectors and implementation of a sustainable control program. Anopheline mosquitoes are routinely identified morphologically, a technique that is time-consuming, needs high level of expertise, and prone to misidentifications especially when considering Amazonian species. The aim of this study was therefore to develop a DNA-based identification technique to supplement traditional morphological identification methods for the discrimination of anopheline mosquitoes collected in French Guiana. The internal transcribed spacer 2 (ITS2) region of ribosomal DNA (rDNA) for anopheline species was amplified by polymerase chain reaction (PCR), and digested with AluI/MspI restriction enzymes. PCR-restriction fragments length polymorphism (RFLP) assay was compared to sequencing of the ITS2 region for validation. Fifteen Anopheles species have shown distinct PCR-RFLP profiles. A concordance of 100% was obtained when identification by PCR-RFLP was compared to sequencing of ITS2. A high throughput, fast, and cost-effective PCR-RFLP assay has been developed for unambiguous discrimination of fifteen anopheline mosquito species from French Guiana including primary and suspected secondary malaria vectors.