The level of embryonation influences detection of Ostertagia ostertagi eggs by semi-quantitative PCR

Markus Drag, Johan Höglund, Peter Nejsum, Stig M Thamsborg, Heidi Enemark

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    Abstract

    The Internal Transcribed Spacer 2 (ITS2) is a candidate diagnostic marker of the pathogenic cattle nematode Ostertagia ostertagi. The aims of this study were: (i) to document and quantify how the development of O. ostertagi eggs affects ITS2 copies under different storage conditions, and (ii) to suggest optimal storage conditions for faecal samples in a diagnostic pipeline that involves detection and semi-quantification by real-time semi-quantitative polymerase chain reaction (qPCR). Eggs of Ostertagia ostertagi were obtained from fresh faeces and stored at 4 °C or 25 °C under aerobic or anaerobic (vacuum packing) conditions. Development was monitored by microscopy for up to 336 h, and the ITS2 copies were determined by qPCR from a fixed number of parasites. Under aerobic conditions at 25 °C, embryonation and a significant increase of ITS2 copies (P 
    Original languageEnglish
    Article number368
    JournalParasites & Vectors
    Volume9
    Issue number1
    Number of pages8
    ISSN1756-3305
    DOIs
    Publication statusPublished - 2016

    Keywords

    • Egg development
    • First-stage larvae
    • ITS2
    • Ostertagia ostertagi
    • Real-time semi-quantitative PCR

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