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PURPOSE:To improve the detection of retinal nerve fiber layer (RNFL) thinning in multiple sclerosis (MS), a special peripapillary ring scanning algorithm (N-site RNFL, N-RNFL) was developed for spectral domain optical coherence tomography (SD-OCT). In contrast to the standard protocol (ST-RNFL) scanning starts nasally, not temporally, and provides an additional sector of analysis, the papillomacular bundle (PMB). We aimed to ascertain whether the temporal RNFL differs between the two techniques, whether N-RNFL is more sensitive than ST-RNFL to detect previous optic neuritis (ON), and whether analyzing the PMB adds additional sensitivity. Furthermore, we investigated whether RNFL is associated with disease severity and/or disease duration. METHODS:We conducted a cross-sectional case-control study of 38 patients with MS, of whom 24 had a history of ON, and 40 healthy controls (HC). Subjects with ON within the previous 6 months were excluded. Records included clinical characteristics, visual evoked potentials (VEP), and SD-OCT in both techniques. RESULTS:In a total of 73 evaluable MS eyes, temporal N-RNFL was abnormal in 17.8%, temporal ST-RNFL in 19.2%, and the PMB-RNFL in 21.9%. In ON eyes, the sensitivity of temporal N-RNFL and ST-RNFL did not differ significantly (37.0%/33.3%, p = 0.556). The sensitivity of VEP was 85.2%. RNFL thickness was associated with disease severity in all eyes, with and without a history of ON, and with disease duration. CONCLUSION:The two OCT techniques detected previous ON with similar sensitivity, but the sensitivity of VEPs was superior to that of both N-RNFL and ST-RNFL. Our results indicate that the widely used ST-RNFL technique is appropriate for peripapillary RNFL measurements in MS patients.