Methods Six experienced glaucoma specialists independently evaluated 50 ONH topographies from 25 adults using HRT software ver. 2.01 in a masked fashion. ONH topographies were obtained from a cohort study of 882 healthy adults and additionally included one patient with glaucomatous eyes. A glaucoma-screening-like setting was intended. The mean interobserver difference was defined as the mean percentual difference between an observer’s analysis and the mean of all six observers for all eyes and all observers. The interobserver range was calculated for each eye as the percentual difference between the lowest and highest measurement, with the highest measurement as denominator. Additionally, Kendall’s coefficient of rank concordance was assessed for the main HRT parameters.

Results Mean disc area ranged from 1.83 ± 0.49 to 2.21 ± 0.40 mm² (mean interobserver difference: 8.3%; interobserver range: 5–50%; rank concordance: 0.86). The lowest mean interobserver differences were found for mean retinal nerve fibre layer thickness (RNFLT; 6.5%), maximum cup depth (2.9%) and cup shape (6.8%). An increased interobserver range was significantly correlated to a low cup to disc area ratio (r = 0.64, P < 0.0001).

Conclusions The observer-dependent diagnostic variability of HRT measurements can lead to divergent diagnostic evaluation of the ONH in a screening setting. Any HRT software relying on a reference database is exposed to relevant observer-related variability of the disc area. For screening purposes, HRT measurements should be completed by other diagnostic methods to compensate for possible diagnostic uncertainty.