1. Discrimination of 15 Amazonian Anopheline Mosquito Species by Polymerase Chain Reaction—Restriction Fragment Length Polymorphism
- Author
-
S B Vezenegho, J Issaly, R Carinci, P Gaborit, R Girod, Isabelle Dusfour, S Briolant, Institut Pasteur de la Guyane, Réseau International des Instituts Pasteur (RIIP), Maladies infectieuses et vecteurs : écologie, génétique, évolution et contrôle (MIVEGEC), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Sud])-Université de Montpellier (UM), Département de Santé Globale - Department Global Health, Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Vecteurs - Infections tropicales et méditerranéennes (VITROME), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut de Recherche Biomédicale des Armées [Brétigny-sur-Orge] (IRBA), Institut Hospitalier Universitaire Méditerranée Infection (IHU Marseille), Institut de Recherche Biomédicale des Armées [Antenne Marseille] (IRBA), Financial support was made available by the French Army (Grant LR607e) for which we are grateful. This work has benefited from an ‘Investissement d’Avenir’ grant managed by Agence Nationale de la Recherche (CEBA, ref. ANR-10-LABX-25-01). This study received a European commission ‘REGPOT-CT-2011-285837-STRonGer’ Grant within the FP7., ANR-10-LABX-0025,CEBA,CEnter of the study of Biodiversity in Amazonia(2010), and European Project: 285837,EC:FP7:REGPOT,FP7-REGPOT-2011-1,STRONGER(2011)
- Subjects
MESH: Malaria ,molecular identification ,Mosquito Vectors ,Polymerase Chain Reaction ,MESH: Anopheles ,malaria vector ,[SDV.MHEP.CSC]Life Sciences [q-bio]/Human health and pathology/Cardiology and cardiovascular system ,[SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases ,DNA, Ribosomal Spacer ,parasitic diseases ,Anopheles ,Animals ,MESH: Animals ,[SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology ,MESH: DNA, Ribosomal Spacer ,[SDV.MHEP.ME]Life Sciences [q-bio]/Human health and pathology/Emerging diseases ,General Veterinary ,MESH: Polymorphism, Restriction Fragment Length ,MESH: Polymerase Chain Reaction ,[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology ,Malaria ,French Guiana ,Infectious Diseases ,Insect Science ,[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,Parasitology ,MESH: Mosquito Vectors ,Polymorphism, Restriction Fragment Length - Abstract
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.
- Published
- 2022