Cécile Nabet1, Abdoulaye K Kone2, Abdoulaye K Dia3, Moussa Sylla4, Magali Gautier5, Mohammed Yattara4, Mahamadou A Thera2, Ousmane Faye3, Leo Braack6,7, Sylvie Manguin8, Abdoul H Beavogui4, Ogobara Doumbo2, Frédérick Gay9, Renaud Piarroux9. 1. Sorbonne Université, INSERM, Institut Pierre-Louis d'Epidémiologie et de Santé Publique, IPLESP, AP-HP, Groupe Hospitalier Pitié-Salpêtrière, Service de Parasitologie-Mycologie, 75013, Paris, France. cecilenabet7@gmail.com. 2. Malaria Research and Training Center (MRTC) UMI 3189 "Environnement, Santé, Sociétés", CNRST, CNRS, UCAD, UGB, University of Sciences, Techniques and Technologies (USTTB), Bamako, Mali. 3. Laboratoire d'Ecologie Vectorielle et Parasitaire (LEVP), Université Cheikh Anta Diop, Dakar, Senegal. 4. Centre de Formation et de Recherche en Santé Rurale de Mafèrinyah, Conakry, Guinea. 5. APHM, Service de Parasitologie-Mycologie, Aix-Marseille Université, Marseille, France. 6. Malaria Consortium, Mahidol Faculty of Tropical Medicine, Bangkok, Thailand. 7. UP Institute for Sustainable Malaria Control, University of Pretoria, Pretoria, South Africa. 8. HydroSciences Montpellier (UMR-HSM), Institut de Recherche pour le Développement (IRD France), CNRS, 34093, Montpellier, France. 9. Sorbonne Université, INSERM, Institut Pierre-Louis d'Epidémiologie et de Santé Publique, IPLESP, AP-HP, Groupe Hospitalier Pitié-Salpêtrière, Service de Parasitologie-Mycologie, 75013, Paris, France.
Abstract
BACKGROUND: Anopheles species identification is essential for an effective malaria vector control programme. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has been developed to identify adult Anopheles species, using the legs or the cephalothorax. The protein repertoire from arthropods can vary according to compartment, but there is no general consensus regarding the anatomic part to be used. METHODS: To determine the body part of the Anopheles mosquitoes best suited for the identification of field specimens, a mass spectral library was generated with head, thorax with wings and legs of Anopheles gambiae, Anopheles arabiensis and Anopheles funestus obtained from reference centres. The MSL was evaluated using two independent panels of 52 and 40 An. gambiae field-collected in Mali and Guinea, respectively. Geographic variability was also tested using the panel from Mali and several databases containing added specimens from Mali and Senegal. RESULTS: Using the head and a database without specimens from the same field collection, the proportion of interpretable and correct identifications was significantly higher than using the other body parts at a threshold value of 1.7 (p < 0.0001). The thorax of engorged specimens was negatively impacted by the blood meal after frozen storage. The addition of specimens from Mali into the database significantly improved the results of Mali panel (p < 0.0001), which became comparable between head and legs. With higher identification scores, the using of the head will allow to decrease the number of technical replicates of protein extract per specimen, which represents a significant improvement for routine use of MALDI-TOF MS. CONCLUSIONS: The using of the head of Anopheles may improve the performance of MALDI-TOF MS. Region-specific mass spectrum databases will have to be produced. Further research is needed to improve the standardization in order to share online spectral databases.
BACKGROUND:Anopheles species identification is essential for an effective malaria vector control programme. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has been developed to identify adult Anopheles species, using the legs or the cephalothorax. The protein repertoire from arthropods can vary according to compartment, but there is no general consensus regarding the anatomic part to be used. METHODS: To determine the body part of the Anopheles mosquitoes best suited for the identification of field specimens, a mass spectral library was generated with head, thorax with wings and legs of Anopheles gambiae, Anopheles arabiensis and Anopheles funestus obtained from reference centres. The MSL was evaluated using two independent panels of 52 and 40 An. gambiae field-collected in Mali and Guinea, respectively. Geographic variability was also tested using the panel from Mali and several databases containing added specimens from Mali and Senegal. RESULTS: Using the head and a database without specimens from the same field collection, the proportion of interpretable and correct identifications was significantly higher than using the other body parts at a threshold value of 1.7 (p < 0.0001). The thorax of engorged specimens was negatively impacted by the blood meal after frozen storage. The addition of specimens from Mali into the database significantly improved the results of Mali panel (p < 0.0001), which became comparable between head and legs. With higher identification scores, the using of the head will allow to decrease the number of technical replicates of protein extract per specimen, which represents a significant improvement for routine use of MALDI-TOF MS. CONCLUSIONS: The using of the head of Anopheles may improve the performance of MALDI-TOF MS. Region-specific mass spectrum databases will have to be produced. Further research is needed to improve the standardization in order to share online spectral databases.
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