DAO 123:101-122 (2017)  -  DOI: https://doi.org/10.3354/dao03093

Diagnostic validation of three test methods for detection of cyprinid herpesvirus 3 (CyHV-3)

Sharon C. Clouthier1,*, Carol McClure2, Tamara Schroeder1, Megan Desai1, Laura Hawley3, Sunita Khatkar1, Melissa Lindsay1, Geoff Lowe3, Jon Richard3, Eric D. Anderson4

1Fisheries and Oceans Canada, Freshwater Institute, 501 University Crescent, Winnipeg, Manitoba R3T 2N6, Canada
2AquaEpi Research, RR3, 300 Heartz Road, Charlottetown, Prince Edward Island C1C 0H8, Canada
3Fisheries and Oceans Canada, Pacific Biological Station, 3190 Hammond Bay Road, Nanaimo, British Columbia V9T 6N7, Canada
4Box 28, Group 30, RR2, Ste Anne, Manitoba R5H 1R2, Canada
*Corresponding author:

ABSTRACT: Cyprinid herpesvirus 3 (CyHV-3) is the aetiological agent of koi herpesvirus disease in koi and common carp. The disease is notifiable to the World Organisation for Animal Health. Three tests—quantitative polymerase chain reaction (qPCR), conventional PCR (cPCR) and virus isolation by cell culture (VI)—were validated to assess their fitness as diagnostic tools for detection of CyHV-3. Test performance metrics of diagnostic accuracy were sensitivity (DSe) and specificity (DSp). Repeatability and reproducibility were measured to assess diagnostic precision. Estimates of test accuracy, in the absence of a gold standard reference test, were generated using latent class models. Test samples originated from wild common carp naturally exposed to CyHV-3 or domesticated koi either virus free or experimentally infected with the virus. Three laboratories in Canada participated in the precision study. Moderate to high repeatability (81 to 99%) and reproducibility (72 to 97%) were observed for the qPCR and cPCR tests. The lack of agreement observed between some of the PCR test pair results was attributed to cross-contamination of samples with CyHV-3 nucleic acid. Accuracy estimates for the PCR tests were 99% for DSe and 93% for DSp. Poor precision was observed for the VI test (4 to 95%). Accuracy estimates for VI/qPCR were 90% for DSe and 88% for DSp. Collectively, the results show that the CyHV-3 qPCR test is a suitable tool for surveillance, presumptive diagnosis and certification of individuals or populations as CyHV-3 free.


KEY WORDS: CyHV-3 · Diagnostic validation · Precision · Accuracy · Quantitative PCR · Virus isolation


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Cite this article as: Clouthier SC, McClure C, Schroeder T, Desai M and others (2017) Diagnostic validation of three test methods for detection of cyprinid herpesvirus 3 (CyHV-3). Dis Aquat Org 123:101-122. https://doi.org/10.3354/dao03093

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