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Published on: May 18, 2020
A multiplex RT-PCR assay for the detection of fish picornaviruses
Sunil K Mor1, Nicholas B D Phelps1, Marisa Barbknecht2
1Minnesota Veterinary Diagnostic Laboratory, Department of Veterinary Population Medicine, University of Minnesota, 1333 Gortner Avenue Street, Paul, MN 55108, USA.
Abstract:
With the emergence of high profile fish diseases in the Great Lakes region, surveillance and regulatory inspections of fish populations have increased. This has resulted in a better understanding of known pathogens and isolation of many new pathogens of fish. In this study, a multiplex RT-PCR assay was developed for the detection of three newly discovered fish picornaviruses: bluegill picornavirus-1 (BGPV-1), fathead minnow picornavirus (FHMPV), and eel picornavirus-1 (EPV-1). This assay was found to be very sensitive with a detection limit of 81.9pg/μl of extracted RNA from a pool of FHMPV and BGPV-1 and was able to detect 501 and 224 gene copies/μl of BGPV-1 and FHMPV, respectively. The assay was highly reproducible and did not cross react with other closely related pathogens. We believe that this new assay provides a rapid and cost effective tool for confirming cell culture isolates and conducting prevalence studies of these newly detected fish picornaviruses.
