Detection of Parapoxvirus in Reindeer

Swab samples from 95 adult reindeer (nasal) and 18 calves (nasal and oral) from 2018 and swab samples from 68 adult reindeer (nasal and oral) from 2019 were analyzed for the presence of parapoxvirus DNA. DNA was extracted from swab samples (Maxwell 16^® Buccal Swab LEV DNA purification kit; Promega, Madison, WI, USA). The PCR reactions were run in a Perkin Elmer GeneAmp^@ PCR System 9700 (Perkin Elmer Corp., Shelton, CT, USA), targeting the putative viral envelope antigen (B2L; primers PPP1 → 5′-gtc gtc cac gag cag ct-3′ and PPP4 → 5′-tac gtg gga agc gcc tcg ct-3′) and a gene encoding a protein inhibiting the granulocyte macrophage colony stimulating factor and interleukin-2 (GIF; primers GIF 5 → 5′-gct cta gga aag atg gcg tg-3′, GIF 6 → 5′-gta ctc ctg gct gaa gag cg -3′), as previously described [46 (link)]. DNA isolated from a skin lesion of a domestic goat (Capra hircus) with CE was used as a positive control (Norwegian Veterinary Institute, Tromsø, Norway). Unused dNTP and primers were removed enzymatically from the PCR amplicons (ExoSAP-IT™; Amersham Pharmacia Biotech, Lund, Sweden) prior to sequencing (BigDye^® Terminator v3.1 cycle sequencing kit; Applied Biosystems, Tønsberg, Norway) in an Applied Biosystems 3130 XL Genetic Analyzer (Applied Biosystems).
DNA was extracted (Maxwell^® 16 LEV Buccal swab DNA kit; Promega, Madison, WI, USA) from leucocytes (“buffy coat”) from two reindeer with antibodies against MCFV and from eight seronegative reindeer. The samples were subjected to a consensus herpesvirus PCR [47 (link)] and an OvHV2 nested PCR [48 (link)] in attempts to amplify herpesvirus-specific or MCFV-specific sequences, respectively.