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

Published online by Cambridge University Press:  19 October 2012

I. C. STEINMANN
Affiliation:
Vector Entomology Unit, Institute of Parasitology, Vetsuisse Faculty, University of Zürich, Zürich, Switzerland
V. PFLÜGER
Affiliation:
Mabritec SA, Riehen, Switzerland
F. SCHAFFNER
Affiliation:
Vector Entomology Unit, Institute of Parasitology, Vetsuisse Faculty, University of Zürich, Zürich, Switzerland
A. MATHIS
Affiliation:
Vector Entomology Unit, Institute of Parasitology, Vetsuisse Faculty, University of Zürich, Zürich, Switzerland
C. KAUFMANN*
Affiliation:
Vector Entomology Unit, Institute of Parasitology, Vetsuisse Faculty, University of Zürich, Zürich, Switzerland
*
*Corresponding author: Vector Entomology Unit, Institute of Parasitology, Vetsuisse Faculty, University of Zürich, Winterthurerstrasse 266a, CH-8057 Zürich, Switzerland. Tel: +41 44 635 85 01. Fax: +41 44 635 89 07. E-mail: ch.kaufmann@access.uzh.ch

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 0·2 mg/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.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2012

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