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Evaluation of matrix-assisted laser desorption/ionization time of flight mass spectrometry for the identification of ceratopogonid and culicid larvae

STEINMANN, I. C. ; PFLÜGER, V. ; SCHAFFNER, F. ; MATHIS, A. ; KAUFMANN, C.

In: Parasitology, 2013, vol. 140, no. 3, p. 318-327

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    Titre
    • Evaluation of matrix-assisted laser desorption/ionization time of flight mass spectrometry for the identification of ceratopogonid and culicid larvae
    Auteur
    • STEINMANN, I. C.. Vector Entomology Unit, Institute of Parasitology, Vetsuisse Faculty, University of Zürich, Zürich, Switzerland
    • PFLÜGER, V.. Mabritec SA, Riehen, Switzerland
    • SCHAFFNER, F.. Vector Entomology Unit, Institute of Parasitology, Vetsuisse Faculty, University of Zürich, Zürich, Switzerland
    • MATHIS, A.. Vector Entomology Unit, Institute of Parasitology, Vetsuisse Faculty, University of Zürich, Zürich, Switzerland
    • KAUFMANN, C.. Vector Entomology Unit, Institute of Parasitology, Vetsuisse Faculty, University of Zürich, Zürich, Switzerland
    Type de document
    • Postprint
    Identifiant OAI-PMH
    • oai:doc.rero.ch:297069
    Summary
    Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) was evaluated for the rapid identification of ceratopogonid larvae. Optimal sample preparation as evaluated with laboratory-reared biting midges Culicoides nubeculosus was the homogenization of gut-less larvae in 10% formic acid, and analysis of 0·2mg/ml crude protein homogenate mixed with SA matrix at a ratio of 1:1·5. Using 5 larvae each of 4 ceratopogonid species (C. nubeculosus, C. obsoletus, C. decor, and Dasyhelea sp.) and of 2 culicid species (Aedes aegypti, Ae. japonicus), biomarker mass sets between 27 and 33 masses were determined. In a validation study, 67 larvae belonging to the target species were correctly identified by automated database-based identification (91%) or manual full comparison (9%). Four specimens of non-target species did not yield identification. As anticipated for holometabolous insects, the biomarker mass sets of adults cannot be used for the identification of larvae, and vice versa, because they share only very few similar masses as shown for C. nubeculosus, C. obsoletus, and Ae. japonicus. Thus, protein profiling by MALDI-TOF as a quick, inexpensive and accurate alternative tool is applicable to identify insect larvae of vector species collected in the field