The primers YHV5f (5′-CGTATTGCATCGAACGTCACTG-3′) and
YHV5r (5′-CAAGATCACTAATAACGCCTGATGC-3′) (De la Rosa-Velez
et al., 2006) were used for the PCR (885 bp), and primers YHV2s (5′-
CGGGGTTACCCGCTTATATT-3′) and YHV2as (5′-GCCTGAGGTGAAGTCC
ATGT-3′), designed as internal sequences, were used for the nested
PCR (400 bp). PCR (total reaction volume=20 μl) and thermal cycling
conditions were as described by De la Rosa-Velez et al. (2006), and 1 μl
of 1:100 diluted PCR product was used in the nested PCR.