DAO 92:1-10 (2010)  -  DOI: https://doi.org/10.3354/dao02277

Development and validation of a TaqMan® PCR assay for the Australian abalone herpes-like virus

Serge Corbeil1,*, Axel Colling1, Lynette M. Williams1, Frank Y. K. Wong2,6, Keith Savin3, Simone Warner2, Bronwyn Murdoch2, Noel O. I. Cogan3, Timothy I. Sawbridge2, Mark Fegan2, Ilhan Mohammad2, Agus Sunarto1, Judith Handlinger4, Stephen Pyecroft4, Marianne Douglas4, Pen H. Chang5, Mark St. J. Crane1

1Australian Animal Health Laboratory, CSIRO Livestock Industries, Geelong, Victoria 3220, Australia
2Biosciences Research Division, Department of Primary Industries, 475 Mickleham Road, Attwood, Victoria 3049, Australia
3Biosciences Research Division, Department of Primary Industries, 1 Park Drive, Bundoora, Victoria 3083, Australia
4Department of Primary Industries, Parks, Water and Environment, 165 Westbury Road, Launceston, Tasmania 7250, Australia
5Department of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, Taipei 10617, Taiwan
6Present address: Australian Animal Health Laboratory, CSIRO Livestock Industries, Geelong, Victoria 3220, Australia

ABSTRACT: The recent emergence of a herpes-like virus in both farmed and wild populations of abalone in Victoria, Australia, has been associated with high mortality rates in animals of all ages. Based on viral genome sequence information, a virus-specific real-time TaqMan assay was developed for detection and identification of the abalone herpes-like virus (AbHV). The assay was shown to be specific as it did not detect other viruses from either the Herpesvirales or the Iridovirales orders which have genome sequence similarities. However, the TaqMan assay was able to detect DNA from the Taiwanese abalone herpes-like virus, suggesting a relationship between the Taiwanese and Australian viruses. In addition, the assay detected <300 copies of recombinant plasmid DNA per reaction. Performance characteristics for the AbHV TaqMan assay were established using 1673 samples from different abalone populations in Victoria and Tasmania. The highest diagnostic sensitivity and specificity were 96.7 (95% CI: 82.7 to 99.4) and 99.7 (95% CI: 99.3 to 99.9), respectively, at a threshold cycle (CT) value of 35.8. The results from 2 separate laboratories indicated good repeatability and reproducibility. This molecular assay has already proven useful in confirming presumptive diagnosis (based on the presence of ganglioneuritis) of diseased abalone in Victorian waters as well as being a tool for surveillance of wild abalone stocks in other parts of Australia.


KEY WORDS: Herpesvirus · Abalone · Real-time TaqMan assay · Diagnosis


Full text in pdf format  
Cite this article as: Corbeil S, Colling A, Williams LM, Wong FYK and others (2010) Development and validation of a TaqMan® PCR assay for the Australian abalone herpes-like virus. Dis Aquat Org 92:1-10. https://doi.org/10.3354/dao02277

Export citation
Mail this link - Contents Mailing Lists - RSS
- -