Kingfisher duo prime platform

Brian tissues were collected from six dead American crows (Corvus brachyrhynchos) between 26 and 28 December 2022 in Prince Edward Island (PEI) and a red fox (Vulpes vulpes) in January 2023 from Newfoundland. The samples were tested in a provincial lab for influenza A viruses and submitted to the National Centre for Foreign Animal Disease (NCFAD), Winnipeg, Manitoba, for confirmatory testing. Total RNA was extracted from all brain tissue samples using the MagMax 1836 Nucleic Acid Isolation Kit using the KingFisher Duo prime platform (ThermoFisher Scientific, Waltham, MA, USA). As in previous studies, influenza A virus nucleic acid was detected initially using the matrix (M) gene and followed by H5 and H7 gene-specific real-time RT-PCR testing [16 (link),17 (link)]. Moreover, viable viruses were isolated in specific pathogen-free embryonated chicken eggs as previously described [18 ].

Total RNA was extracted from clinical specimens (swabs and tissues) and virus isolates using the MagMax 1836 Nucleic Acid Isolation Kit using the KingFisher Duo prime platform (ThermoFisher Scientific, Waltham, MA, USA). The presence of IAV genomic material was verified using the matrix gene-specific real-time RT–PCR followed by H5-specific real-time RT–PCR as described previously [27 (link),28 (link)].