Abstract
BackgroundThe identification of mosquito vectors is typically based on morphological characteristics using morphological keys of determination, which requires entomological expertise and training. The use of protein profiling by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), which is increasingly being used for the routine identification of bacteria, has recently emerged for arthropod identification.MethodsTo investigate the usefulness of MALDI-TOF-MS as a mosquito identification tool, we tested protein extracts made from mosquito legs to create a database of reference spectra. The database included a total of 129 laboratory-reared and field-caught mosquito specimens consisting of 20 species, including 4 Aedes spp., 9 Anopheles spp., 4 Culex spp., Lutzia tigripes, Orthopodomyia reunionensis and Mansonia uniformis. For the validation study, blind tests were performed with 76 specimens consisting of 1 to 4 individuals per species. A cluster analysis was carried out using the MALDI-Biotyper and some spectra from all mosquito species tested.ResultsBiomarker mass sets containing 22 and 43 masses have been detected from 100 specimens of the Anopheles, Aedes and Culex species. By carrying out 3 blind tests, we achieved the identification of mosquito vectors at the species level, including the differentiation of An. gambiae complex, which is possible using MALDI-TOF-MS with 1.8 as the cut-off identification score. A cluster analysis performed with all available mosquito species showed that MALDI-Biotyper can distinguish between specimens at the subspecies level, as demonstrated for An gambiae M and S, but this method cannot yet be considered a reliable tool for the phylogenetic study of mosquito species.ConclusionsWe confirmed that even without any specific expertise, MALDI-TOF-MS profiling of mosquito leg protein extracts can be used for the rapid identification of mosquito vectors. Therefore, MALDI-TOF-MS is an alternative, efficient and inexpensive tool that can accurately identify mosquitoes collected in the field during entomological surveys.
Highlights
Over the last few decades, numerous mosquito-borne infections have emerged or reemerged throughout the world
The MALDI-TOF-MS analysis of the protein extracts prepared from mosquito legs showed spectra with peaks of high intensities in the range of 2– 20 kDa
The 155 peaks detected for the three mosquito genera are higher than the number of peaks obtained for cryptic Anopheles gambiae complex but lower than those obtained for Culicoides biting midges [32]
Summary
Over the last few decades, numerous mosquito-borne infections have emerged or reemerged throughout the world. The medical importance and burden of these infections is enormous [1] They are not limited to tropical areas, where malaria, dengue, and Chikungunya are well known threats to the local population and returning travelers [1,2]. With the growing importance of mosquito-borne diseases, entomological surveys, including the collection and identification of mosquitoes, are needed to better understand transmission dynamics [5] These surveys are essential in the control of vectorborne diseases because they provide information about the vector species involved in the transmission, which is essential for planning effective control measures and monitoring their impact [6]. The use of protein profiling by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), which is increasingly being used for the routine identification of bacteria, has recently emerged for arthropod identification
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