Mean 6 SD G6PD activity for normal samples using the digital microfluidic method and the standard method, respectively, was 9.7 6 2.8 and 11.1 6 3.0 U/g hemoglobin (Hb), respectively; for G6PD-deficient samples, it was 0.8 6 0.7 and 1.4 6 0.9 U/g Hb. Bland-Altman analysis determined a mean difference of –0.96 6 1.8 U/g Hb between the digital microfluidic fluorescence results and the standard biochemical test results. The lower and upper limits for the digital microfluidic platform were 4.5 to 19.5 U/g Hb for normal samples and 0.2 to 3.7 U/g Hb for G6PD-deficient samples. The lower and upper limits for the Stanford method were 5.5 to 20.7 U/g Hb for normal samples and 0.1 to 2.8 U/g Hb for G6PD-deficient samples. The measured activity discriminated between G6PD-deficient samples and normal samples with no overlap.