title: Standardization of sample homogenization for mosquito identification using an innovative proteomic tool based on protein profiling.
abstract: The rapid spread of vector-borne diseases demands the development of an innovative strategy for arthropod monitoring. The emergence of MALDI-TOF MS as a rapid, low-cost, and accurate tool for arthropod identification is revolutionizing medical entomology. However, as MS spectra from an arthropod can vary according to the body part selected, the sample homogenization method used and the mode and duration of sample storage, standardization of protocols is indispensable prior to the creation and sharing of an MS reference spectra database. In the present study, manual grinding of Anopheles gambiae Giles and Aedes albopictus mosquitoes at the adult and larval (L3) developmental stages was compared to automated homogenization. Settings for each homogenizer were optimized, and glass powder was found to be the best sample disruptor based on its ability to create reproducible and intense MS spectra. In addition, the suitability of common arthropod storage conditions for further MALDI-TOF MS analysis was kinetically evaluated. The conditions that best preserved samples for accurate species identification by MALDI-TOF MS were freezing at -20°C or in liquid nitrogen for up to 6 months. The optimized conditions were objectified based on the reproducibility and stability of species-specific MS profiles. The automation and standardization of mosquito sample preparation methods for MALDI-TOF MS analyses will popularize the use of this innovative tool for the rapid identification of arthropods with medical interest.
authors.affiliation: Aix Marseille Université, Unité de Recherche en Maladies Infectieuses et Tropicales Emergentes (URMITE), UM63, CNRS 7278, IRD 198 (Dakar, Sénégal), Inserm 1095, WHO Collaborative Center for Rickettsioses and Other Arthropod-Borne Bacterial Diseases, Faculté de Médecine, Marseille, France.|Laboratoire de Biodiversité et Environnement : Interactions génomes, Faculté des Sciences Biologiques, Université des Sciences et de la Technologie Houari Boumediene (USTHB), Bab Ezzouar, Algiers, Algeria.
authors.forename: Amira
authors.initials: A
authors.lastname: Nebbak
authors.affiliation: Aix Marseille Université, Unité de Recherche en Maladies Infectieuses et Tropicales Emergentes (URMITE), UM63, CNRS 7278, IRD 198 (Dakar, Sénégal), Inserm 1095, WHO Collaborative Center for Rickettsioses and Other Arthropod-Borne Bacterial Diseases, Faculté de Médecine, Marseille, France.|Department of Epidemiology, Gillings School of Global Public Health, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
authors.forename: Alexandra C
authors.initials: AC
authors.lastname: Willcox
authors.affiliation: Aix Marseille Université, Unité de Recherche en Maladies Infectieuses et Tropicales Emergentes (URMITE), UM63, CNRS 7278, IRD 198 (Dakar, Sénégal), Inserm 1095, WHO Collaborative Center for Rickettsioses and Other Arthropod-Borne Bacterial Diseases, Faculté de Médecine, Marseille, France.|Laboratoire de Biodiversité et Environnement : Interactions génomes, Faculté des Sciences Biologiques, Université des Sciences et de la Technologie Houari Boumediene (USTHB), Bab Ezzouar, Algiers, Algeria.
authors.forename: Idir
authors.initials: I
authors.lastname: Bitam
authors.affiliation: Aix Marseille Université, Unité de Recherche en Maladies Infectieuses et Tropicales Emergentes (URMITE), UM63, CNRS 7278, IRD 198 (Dakar, Sénégal), Inserm 1095, WHO Collaborative Center for Rickettsioses and Other Arthropod-Borne Bacterial Diseases, Faculté de Médecine, Marseille, France.
authors.forename: Didier
authors.initials: D
authors.lastname: Raoult
authors.affiliation: Aix Marseille Université, Unité de Recherche en Maladies Infectieuses et Tropicales Emergentes (URMITE), UM63, CNRS 7278, IRD 198 (Dakar, Sénégal), Inserm 1095, WHO Collaborative Center for Rickettsioses and Other Arthropod-Borne Bacterial Diseases, Faculté de Médecine, Marseille, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Parola
authors.affiliation: Aix Marseille Université, Unité de Recherche en Maladies Infectieuses et Tropicales Emergentes (URMITE), UM63, CNRS 7278, IRD 198 (Dakar, Sénégal), Inserm 1095, WHO Collaborative Center for Rickettsioses and Other Arthropod-Borne Bacterial Diseases, Faculté de Médecine, Marseille, France.|Unité Parasitologie et Entomologie, Département des Maladies Infectieuses, Institut de Recherche Biomédicale des Armées, Marseille, France.
authors.forename: Lionel
authors.initials: L
authors.lastname: Almeras
mesh_terms: D000818:Animals; D016000:Cluster Analysis; D009033:Culicidae / Q000737:chemistry* / Q000145:classification; D019476:Insect Proteins / Q000032:analysis*; D007814:Larva / Q000737:chemistry; D040901:Proteomics / Q000191:economics / Q000379:methods* / Q000592:standards; D013045:Species Specificity; D013048:Specimen Handling; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000191:economics / Q000379:methods* / Q000592:standards
chemical_list: D019476:Insect Proteins
keywords: Imago; Innovative strategy; Larva; MALDI-TOF MS; Mosquito identification; Standardized protocol; Technology