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

Serge Corbeil; Axel Colling; Lynette M. Williams; Frank Y. K. Wong; Keith Savin; Simone Warner; Bronwyn Murdoch; Noel O. I. Cogan; Timothy I. Sawbridge; Mark Fegan; Ilhan Mohammad; Agus Sunarto; Judith Handlinger; Stephen Pyecroft; Marianne Douglas; Pen H. Chang; Mark St. J. Crane

doi:10.3354/dao02277

DAO

Diseases of Aquatic Organisms

via Mailchimp

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 Corbeil^1,*, Axel Colling¹, Lynette M. Williams¹, Frank Y. K. Wong^2,6, 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

*Email: serge.corbeil@csiro.au

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 (C_T) 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
Share: Facebook - - linkedIn

Cited by