To assess precision and accuracy of a new objective algorithm using binarization in a software for identifying the hyperautofluorescent ring (AF ring) in retinitis pigmentosa (RP) compared with subjective visual inspection. Ultra-widefield AF images were obtained from 23 eyes of 13 patients with retinitis pigmentosa (RP). We defined the borders of the AF rings using semiautomatic binarization algorithm in Fiji software. We compared the degree of precision (intra- and interrater agreements) of this algorithm and that of subjective visual inspection (freehand method) using Jaccard indices (JIs). To compare the classification performance (whether 68 points of Humphrey Field Analyzer is classified as inside, on, or outside AF rings), we calculated percent agreement and weighted kappa statistic between the two methods. The relationship between the distance from the AF ring and retinal sensitivities was also investigated. The binarization method showed significantly higher JIs than the freehand method (for interrater: 0.94-0.95 vs. 0.73-0.78, respectively, = 0.002; for intrarater: 0.95 vs. 0.68-0.71, respectively, = 0.005). Percent agreement for classification between the two methods were 0.94 and weighted kappa statistic was 0.94 ( < 0.001). The retinal sensitivities decreased significantly and eccentrically from 2° inside to 3° outside the AF ring. Defining the AF ring in RP using the binarization algorithm showed significantly higher precision and the same degree of accuracy compared with visual inspection. This novel method may enable quantitative analysis of the AF ring, an indicator of retinal function in RP.