Agreement between automated and manual quantification of corneal nerve fiber length: Implications for diabetic neuropathy research

Aims: Quantification of corneal nerve fiber length (CNFL) by in vivo corneal confocal microscopy represents a promising diabetic neuropathy biomarker, but applicability is limited by resource-intensive image analysis. We aimed to evaluate, in cross-sectional analysis of non-diabetic controls and patients with type 1 and type 2 diabetes with and without neuropathy, the agreement between manual and automated analysis protocols. Methods: Sixty-eight controls, 139 type 1 diabetes, and 249 type 2 diabetes participants underwent CNFL measurement (N = 456). Neuropathy status was determined by clinical and electrophysiological criteria. CNFL was determined by manual (CNFLmanual, reference standard) and automated (CNFLAuto) protocols, and results were compared for correlation and agreement using Spearman coefficients and the method of Bland and Altman (CNFLManual subtracted from CNFLAuto). Results: Participants demonstrated broad variability in clinical characteristics associated with neuropathy. The mean age, diabetes duration, and HbA1c were 53 +/- 18 years, 15.9 +/- 12.6 years, and 7.4 +/- 1.7%, respectively, and 218 (56%) individuals with diabetes had neuropathy. Mean CNFManual was 15.1 +/- 4.9 mm/mm(2), and mean CNFLAuto was 10.5 +/- 3.7 mm/mm(2) (CNFLAuto underestimation bias, 4.6 +/- 2.6 mm/mm(2) corresponding to 29 17%). Percent bias was similar across non-diabetic controls (-33 +/- 12%), type 1 (-30 +/- 20%), and type 2 diabetes (-28 +/- 16%) subgroups (ANOVA, p = 0.068), and similarly in diabetes participants with and without neuropathy. Levels of CNFLAuto and CNFLmanual were both inversely associated with neuropathy status. Conclusions: Although CNFLAuto substantially underestimated CNFLManual, its bias was non-differential between diverse patient groups and its relationship with neuropathy status was preserved. Determination of diagnostic thresholds specific to CNFLAuto should be pursued in diagnostic studies of diabetic neuropathy. (C) 2016 Elsevier Inc. All rights reserved.