Screening for Equine Parvovirus-H Variants

We aligned the nonstructural (NS) protein and virion protein (VP) of EqPV-H to all known parvovirus proteins. We used nucleotide and amino acid motifs showing relative conservation among different virus lineages to make primers for screening samples for EqPV-H and related variants. All PCR mixtures used AmpliTaq Gold 360 master mix (catalog no. 4398881; Applied Biosystems, Foster City, CA, USA) and 2 μL of extracted nucleic acids. The EqPV-H NS gene PCR used primer pair EqPV ak1 (5′-GGAGAAGAGCGCAACAAATGCA-3′) and EqPV ak2 (5′-AAGACATTTCCGGCCGTGAC-3′) in the first round of PCR and the pair EqPV ak3 (5′-GCGCAACAAATGCAGCGGTTCGA-3′) and EqPV ak4 (5′-GGCCGTGACGACGGTGATATC-3′) in the second round of PCR. The EqPV-H VP gene PCR used primer pair EqPV ak5 (5′-GTCGCTGCATTCTGAGTCC-3′) and EqPV ak6 (5′-TGGGATTATACTGTCTACGGGT-3′) in the first round of PCR and the pair EqPV ak7 (5′-CTGCATTCTGAGTCCGTGGCC-3′) and EqPV ak8 (5′-CTGTCTACGGGTATCCCATACGTA-3′) in the second round of PCR.