Figure 1
Establishment of real-time PCR. A. Standard curves (based on plasmid DNA) indicating the linearity and efficiency for detecting NS1 by real-time PCR. The x-axis represents copies of plasmid DNA in 10-fold dilutions, and the y-axis represents the fluorescence data used for cycle threshold (Ct) determinations. The assays were linear in the range of 109 to 102 template copies/μL, with an R2 of 0.996 and a reaction efficiency of 100% for NS1. B. The results of the Quantitation data for Cycling A. FAM. The amplification product was about 123 bp long, and no false amplification was observed. The detection limit of the real-time PCR for the NS1 gene of PPV was 1.00 × 102 copies/μL. C. Sensitivity of normal PCR. N: Negative Control. M: DNA marker DL2000. Lane 1-9: The standard plasmid was 10-fold serially diluted as template. D. Specificity of the real-time PCR assay. PPV:Positive sample; A-F:Negative control, PCV2, PRV, PRRSV, CFSV, JEV, and H2O control.