Optimized PCR assay for detection of white spot syndrome virus (WSSV)

Linda M. Nunan, Donald V. Lightner

    Research output: Contribution to journalArticlepeer-review

    65 Scopus citations

    Abstract

    A rapid PCR assay for detection of white spot syndrome virus (WSSV) was developed based on the nested PCR procedure described by Lo et al. (1996) and outlined as the recommended PCR diagnostic assay in the Manual of Diagnostic Tests for Aquatic Animals published by the Office of International Epizootics (OIE, 2009). The optimized procedure incorporated the second step primers used in the nested WSSV PCR. By adjusting the annealing temperature and shortening the cycling times, this modified assay is substantially faster and as sensitive as the recommended OIE protocol. The modified PCR test was compared directly to the two-step nested PCR protocol and a modified nested procedure. The sensitivity of the published assay was determined by template dilutions of semi-purified WSSV virions that had been quantitated using real-time PCR for detection of WSSV. Various isolates were tested using the modified procedure, to ensure that the assay was able to detect WSSV from different geographical locations.

    Original languageEnglish (US)
    Pages (from-to)318-321
    Number of pages4
    JournalJournal of Virological Methods
    Volume171
    Issue number1
    DOIs
    StatePublished - Jan 2011

    Keywords

    • PCR
    • Peneaid shrimp
    • White spot syndrome virus (WSSV)

    ASJC Scopus subject areas

    • Virology

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