A multiplex RT-PCR assay for the detection of fish picornaviruses

Sunil K. Mor, Nicholas B.D. Phelps, Marisa Barbknecht, Michael A. Hoffman, Sagar M. Goyal

Research output: Contribution to journalArticle

2 Citations (Scopus)

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 81.9. pg/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.

Original languageEnglish (US)
Pages (from-to)131-134
Number of pages4
JournalJournal of Virological Methods
Volume221
DOIs
StatePublished - Jan 1 2015

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Picornaviridae
Multiplex Polymerase Chain Reaction
Fishes
Perciformes
Cyprinidae
Great Lakes Region
Fish Diseases
Eels
Limit of Detection
Cell Culture Techniques
Cross-Sectional Studies
RNA
Costs and Cost Analysis

Keywords

  • Bluegill picornavirus
  • Eel picornavirus
  • Fathead minnow picornavirus
  • Multiplex RT-PCR
  • Reproducibility
  • Sensitivity
  • Specificity

Cite this

A multiplex RT-PCR assay for the detection of fish picornaviruses. / Mor, Sunil K.; Phelps, Nicholas B.D.; Barbknecht, Marisa; Hoffman, Michael A.; Goyal, Sagar M.

In: Journal of Virological Methods, Vol. 221, 01.01.2015, p. 131-134.

Research output: Contribution to journalArticle

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AU - Goyal, Sagar M.

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