A new tool for the molecular identification of Culicoides species of the Obsoletus group: the glass slide microarray approach

I Deblauwe, JC De Witte, G De Deken, R De Deken, M Madder, S Van Erk, FA Hoza, D Lathouwers, D Geysen

    Research output: Contribution to journalA1: Web of Science-articlepeer-review

    Abstract

    Culicoides species of the Obsoletus group (Diptera: Ceratopogonidae) are potential vectors of bluetongue virus serotype 8 (BTV 8), which was introduced into central Western Europe in 2006. Correct morphological species identification of Obsoletus group females is especially difficult and molecular identification is the method of choice. In this study we present a new molecular tool based on probe hybridization using a DNA microarray format to identify Culicoides species of the Obsoletus group. The internal transcribed spacer 1 (ITS1) gene sequences of 55 Culicoides belonging to 13 different species were determined and used, together with 19 Culicoides ITS1 sequences sourced from GenBank, to design species-specific probes for the microarray test. This test was evaluated using the amplified ITS1 sequences of another 85 Culicoides specimens, belonging to 11 species. The microarray test successfully identified all samples (100%) of the Obsoletus group, identifying each specimen to species level within the group. This test has several advantages over existing polymerase chain reaction (PCR)-based molecular tools, including possible capability for parallel analysis of many species, high sensitivity and specificity, and low background signal noise. Hand-spotting of the microarray slide and the use of detection chemistry make this alternative technique affordable and feasible for any diagnostic laboratory with PCR facilities.
    Original languageEnglish
    JournalMedical and Veterinary Entomology
    Volume26
    Issue number1
    Pages (from-to)83-91
    ISSN0269-283X
    DOIs
    Publication statusPublished - 2012

    Keywords

    • B780-tropical-medicine
    • Viral diseases
    • Animal diseases
    • Bluetongue
    • Vectors
    • Mosquitoes
    • Culicoides
    • Obsoletus
    • Molecular
    • Identification
    • DNA microarrays
    • Hybridization
    • Internal transcribed spacer 1
    • ITS1
    • Gene sequencing
    • Affordability
    • Feasibility
    • Laboratory techniques and procedures

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