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

作者：Mor Sunil K^*; Phelps Nicholas B D; Barbknecht Marisa; Hoffman Michael A; Goyal Sagar M

来源：Journal of Virological Methods, 2015, 221: 131-134.

DOI：10.1016/j.jviromet.2015.04.025

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

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/mu l of extracted RNA from a pool of FHMPV and BGPV-1 and was able to detect 501 and 224 gene copies/mu l 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.

• 出版日期2015-9-1