Fig. 1. Simultaneous detection using the finally selected primer combinations for the three viruses (Clover yellow vein virus, ClYVV; Peanut mottle virus, PeMoV, and Tomato spotted wilt virus, TSWV) by multiplex PCR. Lane M: 1kb DNA ladder (BioFACT, Daejeon, Korea), Lane 1 and 8: 447 bp for ClYYV; Lane 2 and 9: 599 and 560 bp for PeMoV; Lane 3 and 10: 947 and 682 bp for TSWV; Lane 4 and 11: ClYVV+PeMoV; Lane 5 and 12: ClYVV+TSWV; Lane 6 and 13: PeMoV+TSWV; Lane 7 and 14: ClYVV+PeMoV+TSWV.
Fig. 2. Sensitivity test of uniplex-, duplex- and multiplex-PCR for the three legume viruses. (A) uniplex-PCR test results of ClYYV, PeMoV, and TSWV using the first primer combination, (B) duplex-PCR test results of ClYVV+PeMoV, ClYVV+TSWV, and PeMoV+TSWV using the first combination, (C) multiplex-PCR test results of ClYVV+PeMoV+TSWV and comparison of the two different primer combination sets no. 1 (left) and 2 (right). Lane M: 1kb DNA ladder (BioFACT, Daejeon, Korea), Lane 1 to 8: 10-1 to 10-8 diluted cDNA.
Table 1. Oligonucleotide sequences for the two different primer combinations used for the detection of the three viruses
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