Sensitivity and specificity plots for real-time AS-PCR assay. (A) Δ reaction curve for real time AS-PCR assay. The MYD88 L265P mutant DNA (OCI-LY3) was diluted with the wild-type DNA (OCI-LY19) at the concentrations indicated in the amplification plot, with the MYD88 L265P allele detectable to a dilution of 0.08%. (B) Standard curve for real time AS-PCR assay. The correlation coefficient was 0.998 with a slope value of −3.48. (C) Dissociation curve for real time AS-PCR assay. Melting curve analysis revealed that the MYD88 L265P mutant-specific amplicon melted at 84°C. A minor nonspecific amplification was only found in the dilution of 0.4% or lower with a melting peak at 80°C. WT, wild-type.