Identification of Neotropical Culex Mosquitoes by MALDI-TOF MS Profiling

Costa, Monique Melo; Guidez, Amandine; Briolant, Sébastien; Talaga, Stanislas; Issaly, Jean; Naroua, Halima; Carinci, Romuald; Gaborit, Pascal; Lee, Anne; Dusfour, Isabelle; Duchemin, Jean‐Bernard; Almeras, Lionel

doi:10.3390/tropicalmed8030168

Cited by 6 publications

(21 citation statements)

References 79 publications

(107 reference statements)

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“…Other approaches for mosquito species identification are currently being considered. Mosquito protein profiling using MALDI-TOF technology is promising [33] , even though it requires a substantial investment in terms of equipment and the databases are still under development [34] . Nowadays, increasingly powerful software solutions make it possible to perform morphometry of mosquito wings [35] and the use of artificial intelligence may allow for automatic visual identification of mosquitoes [36] .…”

Section: Introductionmentioning

confidence: 99%

A novel mosquito species identification method based on PCR and capillary electrophoresis

Chabanol

Romoli

Talaga

et al. 2023

Preprint

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In the Anopheles genus, various mosquito species are able to transmit Plasmodium parasites responsible for malaria, while others are non-vectors. In an effort to better understand the biology of Anopheles species and to quantify transmission risk in an area, the identification of mosquito species collected on the field is an essential but problematic task. Morphological identification requires expertise, well-preserved specimens and high-quality equipment, and it does not allow any subsequent verification when samples are later used in a destructive treatment. Moreover, it involves physical manipulations that are not compatible with experiments requiring fast sampling and processing of specimens, hence species identification is often based on DNA sequencing of reference genes or region such as the Internal Transcribed Spacer 2 (ITS2) region of nuclear ribosomal DNA. Sequencing ITS2 for numerous samples is costly, but the design of species-specific PCR primers is not always possible when local species diversity is high. Here, we introduce a molecular technique of species identification based on precise determination of ITS2 length combined with a simple visual observation, the color of mosquito hindleg tip. DNA extracted from field-collected Anopheles mosquitoes was amplified with universal Anopheles ITS2 primers and analyzed with a capillary electrophoresis device, which precisely determines the size of the fragments. We defined windows of amplicon sizes combined with fifth hind tarsus color, which allow to discriminate the major Anopheles species found in our collections. We validated our parameters via Sanger sequencing of the ITS2 amplicons. This method can be particularly useful in situations with a moderate species diversity, i.e. when the number of local species is too high to define species-specific primers but low enough to avoid individual ITS2 sequencing. This tool will be of interest to evaluate local malaria transmission risk and this approach may further be implemented for other mosquito genera.

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Section: Introductionmentioning

confidence: 99%

A novel mosquito species identification method based on PCR and capillary electrophoresis

Chabanol

Romoli

Talaga

et al. 2023

Preprint

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“…LSVs ranging from 0 to 3 were obtained for each spectrum of the samples tested. According to previous studies [28,37], LSVs greater than 1.8 were considered reliable for species identification. For evaluating it performance, the sensitivity, the specificity, the accuracy and the Cohen's κ coefficient, corresponding to the degree of agreement [38], were calculated.…”

Section: Database Creation and Blind Testsmentioning

confidence: 96%

Potential of MALDI-TOF MS biotyping to detect deltamethrin resistance in the dengue vector Aedes aegypti

Almeras,

Costa,

Amalvict

et al. 2024

PLoS ONE

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Insecticide resistance in mosquitoes is spreading worldwide and represents a growing threat to vector control. Insecticide resistance is caused by different mechanisms including higher metabolic detoxication, target-site modification, reduced penetration and behavioral changes that are not easily detectable with simple diagnostic methods. Indeed, most molecular resistance diagnostic tools are costly and labor intensive and then difficult to use for routine monitoring of insecticide resistance. The present study aims to determine whether mosquito susceptibility status against the pyrethroid insecticides (mostly used for mosquito control) could be established by the protein signatures of legs and/or thoraxes submitted to MALDI-TOF Mass Spectrometry (MS). The quality of MS spectra for both body parts was controlled to avoid any bias due to unconformity protein profiling. The comparison of MS profiles from three inbreeds Ae. aegypti lines from French Guiana (IRF, IR03, IR13), with distinct deltamethrin resistance genotype / phenotype and the susceptible reference laboratory line BORA (French Polynesia), showed different protein signatures. On both body parts, the analysis of whole protein profiles revealed a singularity of BORA line compared to the three inbreeding lines from French Guiana origin, suggesting that the first criteria of differentiation is the geographical origin and/or the breeding history rather than the insecticide susceptibility profile. However, a deeper analysis of the protein profiles allowed to identify 10 and 11 discriminating peaks from leg and thorax spectra, respectively. Among them, a specific peak around 4870 Da was detected in legs and thoraxes of pyrethroid resistant lines compared to the susceptible counterparts hence suggesting that MS profiling may be promising to rapidly distinguish resistant and susceptible phenotypes. Further work is needed to confirm the nature of this peak as a deltamethrin resistant marker and to validate the routine use of MS profiling to track insecticide resistance in Ae. aegypti field populations.

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“…This second dataset included MS spectra from Neotropical Culex mosquitoes (Diptera: Culicidae) coming from a work recently published [31]. The methods and protocols used for mosquito collection, for their morphological and/or molecular identification, for sample preparation and MS submission were detailed in this previous study [31]. Briefly, this dataset was composed of MS spectra from legs and thoraxes of 13 distinct Culex species collected in the field, in French Guiana.…”

Section: Use Case N°2: Culex Genusmentioning

confidence: 99%

MSProfileR: An Open-Source Software for Quality Control of MALDI-TOF Spectra

Ben Hamouda,

Estellon,

Himet

et al. 2024

Preprint

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In the early 2000s, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) emerged as a performant and relevant tool for identifying micro-organisms. Since then, it has become practically essential for identifying bacteria in microbiological diagnostic laboratories. In the last decade, it was successfully applied for arthropod identification allowing to distinguish vectors from non-vectors of infectious diseases. However, identification failures are not rare, hampering it widely use. Failure is generally attributed either to the absence of respective counter species MS spectra in the database, or to the insufficient quality of query MS spectra (i.e. lower intensity and diversity of MS peaks detected). To avoid matching errors due to non-compliant spectra, the development of a strategy for detecting and excluding outlier MS profiles became compulsory. To this end, we created MSProfileR, an R package leading to a bioinformatics tool through a simple installation, integrating the control quality system of MS spectra and an analysis pipeline including peak detection and MS spectra comparisons. MSProfileR can also add metadata concerning the sample that the spectra are derived from. MSProfileR has been developed in the R environment and offers a user-friendly web interface using the R Shiny framework. It is available on Microsoft Windows as a web browser application by simply navigation using the link of the package on the Github. MSProfileR is therefore accessible to non-computer specialists and is freely available to the scientific community. We evaluated MSProfileR using two datasets including exclusively MS spectra from arthropods. In addition to coherent sample classification, outlier MS spectra were detected in each dataset confirming the value of MSProfileR.

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Identification of Neotropical Culex Mosquitoes by MALDI-TOF MS Profiling

Cited by 6 publications

References 79 publications

A novel mosquito species identification method based on PCR and capillary electrophoresis

A novel mosquito species identification method based on PCR and capillary electrophoresis

Potential of MALDI-TOF MS biotyping to detect deltamethrin resistance in the dengue vector Aedes aegypti

MSProfileR: An Open-Source Software for Quality Control of MALDI-TOF Spectra

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