title: Identification of phlebotomine sand flies (Diptera: Psychodidae) by matrix-assisted laser desorption/ionization time of flight mass spectrometry.
abstract: BACKGROUND
  Phlebotomine sand flies are incriminated in the transmission of several human and veterinary pathogens. To elucidate their role as vectors, proper species identification is crucial. Since traditional morphological determination is based on minute and often dubious characteristics on their head and genitalia, which require certain expertise and may be damaged in the field-collected material, there is a demand for rapid, simple and cost-effective molecular approaches.
  
  
  METHODS
  Six laboratory-reared colonies of phlebotomine sand flies belonging to five species and four subgenera (Phlebotomus, Paraphlebotomus, Larroussius, Adlerius) were used to evaluate the discriminatory power of matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). Various storage conditions and treatments, including the homogenization in either distilled water or given concentrations of formic acid, were tested on samples of both sexes.
  
  
  RESULTS
  Specimens of all five analysed sand fly species produced informative, reproducible and species-specific protein spectra that enabled their conclusive species identification. The method also distinguished between two P. sergenti colonies originating from different geographical localities. Protein profiles within a species were similar for specimens of both sexes. Tested conditions of specimen storage and sample preparation give ground to a standard protocol that is generally applicable on analyzed sand fly specimens.
  
  
  CONCLUSIONS
  Species identification of sand flies by MALDI-TOF MS is feasible and represents a novel promising tool to improve biological and epidemiological studies on these medically important insects.
authors.affiliation: Department of Parasitology, Faculty of Science, Charles University in Prague, Prague, Czech Republic. icejumper@seznam.cz.
authors.forename: Vit
authors.initials: V
authors.lastname: Dvorak
authors.forename: Petr
authors.initials: P
authors.lastname: Halada
authors.forename: Kristyna
authors.initials: K
authors.lastname: Hlavackova
authors.forename: Emmanouil
authors.initials: E
authors.lastname: Dokianakis
authors.forename: Maria
authors.initials: M
authors.lastname: Antoniou
authors.forename: Petr
authors.initials: P
authors.lastname: Volf
mesh_terms: D000818:Animals; D016000:Cluster Analysis; D005260:Female; D019476:Insect Proteins / Q000378:metabolism*; D008297:Male; D040901:Proteomics; D011576:Psychodidae / Q000145:classification* / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D019476:Insect Proteins