PCR assay for discriminating between infectious hypodermal and hematopoietic necrosis virus (IHHNV) and virus-related sequences in the genome of Penaeus monodon

Kathy F.J. Tang, Solangel A. Navarro, Donald V. Lightner

    Research output: Contribution to journalArticlepeer-review

    78 Scopus citations

    Abstract

    We developed a PCR assay that can detect infectious hypodermal and hematopoietic necrosis virus (IHHNV) but that does not react with IHHNV-related sequences in the genome of Penaeus monodon from Africa and Australia. IHHNV is a single-stranded DNA virus that has caused severe mortality and stunted growth in penaeid shrimp. Recently, IHHNV-related sequences were found in the genome of some stocks of P. monodon from Africa and Australia. These virus-related sequences have a high degree of similarity (86 and 92 % identities in nucleotide sequence) to the viral genome, which has often generated false-positive reactions during PCR screening of these stocks. For this assay, a pair of IHHNV primers (IHHNV309F/R) was selected. The sequences of these primers match (100% of nucleotides) the target sequence in IHHNV, but mismatch 9 or 12 nucleotides of the genomic IHHNV-related sequences. This PCR assay was tested with various IHHNV isolates and with a number of samples of shrimp DNA that contained IHHNV-related sequences. This assay can reliably distinguish IHHNV DNA from shrimp DNA: it only detects IHHNV. Also, this pair of primers was included in a duplex PCR to detect IHHNV and simultaneously determine the presence of an IHHNV-related sequence. Using these primers, the PCR assay has a sensitivity equivalent to a PCR assay commonly used for detecting IHHNV in Litopenaeus vannamei, and can be used for routine detection.

    Original languageEnglish (US)
    Pages (from-to)165-170
    Number of pages6
    JournalDiseases of aquatic organisms
    Volume74
    Issue number2
    DOIs
    StatePublished - Feb 28 2007

    Keywords

    • IHHNV-related sequences
    • Infectious hypodermal and hematopoietic necrosis virus (IHHNV)
    • PCR
    • Penaeus monodon

    ASJC Scopus subject areas

    • Ecology, Evolution, Behavior and Systematics
    • Aquatic Science

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