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Diseases of Aquatic Organisms

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DAO 69:137-143 (2006)  -  doi:10.3354/dao069137

Detection of the herpesviral hematopoietic necrosis disease agent (Cyprinid herpesvirus 2) in moribund and healthy goldfish: validation of a quantitative PCR diagnostic method

A. E. Goodwin*, G. E. Merry, J. Sadler

University of Arkansas at Pine Bluff, Aquaculture/Fisheries Center, 1200 N. University Dr., Mail Slot 4912, Pine Bluff, Arkansas 71601, USA

ABSTRACT: Cyprinid herpesvirus 2 (CyHV-2) is a pathogen of goldfish Carassius auratus auratus L. that causes herpesviral hematopoietic necrosis (HVHN) disease. The disease is associated with necrosis of hematopoietic tissues and anemia with high mortality. We have developed a real time 5’-nuclease PCR method (Taqman) that quantitatively detects CyHV-2 with a linear response over 8 logs of target concentration. The coefficient of variability on replicate samples tested on different days was 13% and the calculated sensitivity approached 1 target molecule per reaction. The assay does not cross-react with other similar fish herpesviruses, including CyHV-1 (carp pox) and CyHV-3 (koi herpesvirus), but reliably detects known CyHV-2 positive fish. The assay detects CyHV-2 not just in clinical cases of HVHN but also in apparently healthy 1 yr old goldfish fingerlings and even in 3 to 5 yr old broodfish.

KEY WORDS: Herpes · Virus · Goldfish · Quantitative PCR · CyHV-2

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