Fig. 4

Sensitivity and accuracy of A2063G and A2064G. A-B: The sensitivity of A2063G and A2064G. Transverse line: The cut-off value line of each mutation. The cut-off value was defined as the mean Ct value plus three standard deviations (SD) of 12 independent determinations of 10⁵ copies wild-type template with mutant-specific primer. The Ct values of the mixtures with mutant template ranging from 0.01 to 100% were less than the cut-off values of detecting A2063G and A2064G assays. These indicated that resistance-associated mutations could still be detected until their proportion was about 0.01%. C-D: The accuracy of A2063G and A2064G. Measured proportion = Measured mutant copy number/ measured total DNA copy number× 100%. Measured and nominal proportions were comparable. The measured proportions of detecting A2063G and A2064G assays were accurate down to 0.1%