Development of a one-step multiplex qRT–PCR assay for the detection of African swine fever virus, classical swine fever virus and atypical porcine pestivirus

Figures

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  Fig. 1. Standard curves of the multiplex qRT-PCR. The triplicate standard curves indicated a linear correlation between the logarithm of the copy number and the Ct values. The concentrations of the standard plasmids (p-ASFV, p-CSFV and p-APPV) ranged from 2.52 × 108 to 2.52 × 100 copies/μL (5.04 × 108 to 5.04 × 100 copies per reaction)

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  Fig. 2. Specificity analysis of the multiplex qRT-PCR using different viral strains. The standard recombinant plasmids (p-ASFV, p-CSFV and p-APPV), ASFV, CSFV, APPV and other viruses (PCV2, PRV, PRRSV, FMDV, PPV, PEDV, TGEV, PRoV, PDCoV, BVDV-1, BVDV-2 and BDV) were used to test the specificity

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  Fig. 3. Sensitivity analysis of the multiplex qRT-PCR. The standard recombinant plasmids (p-ASFV, p-CSFV and p-APPV) were used to test the sensitivity. 1-9: 2.52 × 107 - 2.52 × 10− 1 copies/μL (final reaction concentrations)

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  Fig. 4. Phylogenetic tree based on partial p72 gene of ASFV. The tree was constructed using the neighbour-joining algorithm of MEGA5.0, and 1000 bootstrap replicates were performed to assign confidences to the groupings. The strains from this study were marked with triangle (△)