Figure legends
Figure 1
LOD of CaPV, PaPV and FMDV PACs by singleplex and multiplex assays. A, CaPV PAC serially diluted in nuclease-free water (1-plex) or in negative skin DNA in the presence (3-plex) or absence (2-plex) of PaPV qPCR primers and probe; B, FMDV PAC (reverse transcribed RNA) serially diluted in nuclease-free water (1-plex) or in negative skin DNA in the presence (3-plex) or absence (2-plex) of PaPV qPCR primers and probe; C, PaPV PAC serially diluted in nuclease-free water (1-plex) or in negative skin DNA in the absence (2-plex) or presence of CaPV qPCR (PaP1, 3-plex) or FMDV RT-qPCR (PaP2, 3-plex) primers and probe. Amplification ofACTB shown in the graph was taken from 3-plex assays only.
Figure 2
LOD of LSDV, GPV, SPV and PaPV by singleplex (1-plex) and CaP rule-out (3-plex) assays. The viral DNA was serially diluted in nuclease-free water (1-plex) or negative skin DNA (3-plex) and used as template. A, serial dilution of LSDV (1-plex) or combined LSDV/PaPV (3-plex); B, serial dilutions of GPV (1-plex) or combined GPV/PaPV (3-plex); C, serial dilutions of SPV (1-plex) or combined SPV/PaPV (3-plex). Not shown in the figure is the LOD of PaPV DNA by singleplex assays which remained same (106 dilution) as the corresponding CaP rule-out assays.
Sources of the viral DNAs: LSDV (LSDV-Uganda), GPV (GPV-Pendik) and SPV (SPV-HELD) DNAs were extracted from CC grown viruses (Supplemental Table 1). The PaPV (BPSV) DNA was extracted from CS of a naturally infected cattle (FADDL/DSS accession ID 15-020595).
Figure 3
LOD of PaPV and the seven serotypes of FMDV by singleplex (1-plex) and FMD rule-out (3-plex) assays. The viral DNA/RNA was serially diluted in nuclease-free water (1-plex) or negative skin DNA (3-plex) and used as template. A, serial dilution of FMDV-A (1-plex) or combined PaPV/FMDV-A (3-plex); B, serial dilution of FMDV-Asia1 (1-plex) or combined PaPV/FMDV-Asia1 (3-plex); C, serial dilution of FMDV-C (1-plex) or combined PaPV/FMDV-C (3-plex); D, serial dilution of FMDV-O (1-plex) or combined PaPV/FMDV-O (3-plex); E, serial dilution of FMDV-SAT1 (1-plex) or combined PaPV/FMDV-SAT1 (3-plex); F, serial dilution of FMDV-SAT2 (1-plex) or combined PaPV/FMDV-SAT2 (3-plex); G, serial dilution of FMDV-SAT3 (1-plex) or combined PaPV/FMDV-SAT3 (3-plex). Not shown in the figure is the LOD of PaPV DNA by singleplex RT-qPCR assays which remained same (106 dilution) as the corresponding FMD rule-out (3-plex) assays.
Sources of the viral DNA/RNA: FMDV RNAs of the serotypes A (A23 Kenya), Asia1 (PAK 1/54), C (C3 RESENDE), O (O1 Manisa), SAT1 (Rhodesia), SAT2 (Zimbabwe 5/81) and SAT3 (South Africa) were extracted from CC grown viruses (Supplemental Table 1). The source of the PaPV DNA was the same as described in the legends to Figure 2.
Supplemental Figure 1
LOD of CaPV, PaPV and FMDV by CaP/FMD rule-out assay. A, combined LSDV/PaPV/FMDV-A (DNA/RNA) serially diluted in negative skin DNA; B combined LSDV/PaPV/FMDV-C serially diluted in negative skin DNA; C. combined LSDV/PaPV/FMDV-O serially diluted in negative skin DNA.
Supplemental Figure 2
LOD of CaPV, PaPV and FMDV by CaP/FMD rule-out assay. A, combined GPV/ PaPV/FMDV-C (DNA/RNA) serially diluted in negative skin DNA; B, combined GPV/PaPV/FMDV-O (DNA/RNA) serially diluted in negative skin DNA; C, combined SPV/ PaPV/FMDV-C (DNA/RNA) serially diluted in negative skin DNA; D, combined SPV/PaPV/FMDV-O (DNA/RNA) serially diluted in negative skin DNA.