ORCID
- Puschendorf, Robert: 0000-0002-8105-171X
- Knight, Mairi: 0000-0001-9388-4850
Abstract
Ranaviruses are emerging pathogens that can cause morbidity, mortality and population declines in ectothermic hosts; however, there is no standardized approach to diagnostics. Here, we compared the inter-assay variation and intra-assay precision among 2 commonly used quantitative PCRs (qPCRs), a conventional and a nested PCR assay (used as a gold standard), using laboratory-propagated ranavirus (FV3 and CMTV) and field-collected samples. A qPCR assay (‘Leung’) detected viral DNA in dilutions 2 orders of magnitude lower than other assays regardless of the viral lineage of the cultured isolate (FV3/CMTV). The second qPCR (‘Brunner’) was slightly more sensitive than the conventional PCR (‘Mao’ assay). For field samples, the Leung qPCR detected all known positives, while the Mao assay PCR only detected 2.5% of the positive samples. Amplicon sequences from the 2 conventional PCRs were shown to be useful for inferring viral lineage. Inaccurate results will bias estimates of the distribution and prevalence of ranaviruses, and together these findings emphasize that molecular assays should be chosen carefully in the context of study aims.
DOI
10.3354/dao03518
Publication Date
2020-09-24
Publication Title
Diseases of Aquatic Organisms
Volume
141
ISSN
0177-5103
Embargo Period
2022-01-21
Organisational Unit
School of Biological and Marine Sciences
First Page
139
Last Page
147
Recommended Citation
Wynne, F. J., Puschendorf, R., Knight, M., & Price, S. (2020) 'Choice of molecular assay determines ranavirus detection probability and inferences about prevalence and occurrence', Diseases of Aquatic Organisms, 141, pp. 139-147. Available at: https://doi.org/10.3354/dao03518
