title: Development and validation of a TaqMan PCR assay for the Australian abalone herpes-like virus.
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.
authors.affiliation: Australian Animal Health Laboratory, CSIRO Livestock Industries, Geelong, Victoria 3220, Australia. serge.corbeil@csiro.au
authors.forename: Serge
authors.initials: S
authors.lastname: Corbeil
authors.forename: Axel
authors.initials: A
authors.lastname: Colling
authors.forename: Lynette M
authors.initials: LM
authors.lastname: Williams
authors.forename: Frank Y K
authors.initials: FY
authors.lastname: Wong
authors.forename: Keith
authors.initials: K
authors.lastname: Savin
authors.forename: Simone
authors.initials: S
authors.lastname: Warner
authors.forename: Bronwyn
authors.initials: B
authors.lastname: Murdoch
authors.forename: Noel O I
authors.initials: NO
authors.lastname: Cogan
authors.forename: Timothy I
authors.initials: TI
authors.lastname: Sawbridge
authors.forename: Mark
authors.initials: M
authors.lastname: Fegan
authors.forename: Ilhan
authors.initials: I
authors.lastname: Mohammad
authors.forename: Agus
authors.initials: A
authors.lastname: Sunarto
authors.forename: Judith
authors.initials: J
authors.lastname: Handlinger
authors.forename: Stephen
authors.initials: S
authors.lastname: Pyecroft
authors.forename: Marianne
authors.initials: M
authors.lastname: Douglas
authors.forename: Pen H
authors.initials: PH
authors.lastname: Changs
authors.forename: Mark St J
authors.initials: MS
authors.lastname: Crane
mesh_terms: D000818:Animals; D001315:Australia; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D006564:Herpesviridae / Q000302:isolation & purification*; D008974:Mollusca / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral