Diabetic autonomic neuropathy (DAN) is a very frequent complication in the diabetic population (type 1 and type 2 diabetes), and patients may suffer debilitating symptoms from various organ systems. In the less symptomatic and even in the asymptomatic condition it severely impacts health. Testing for DAN is currently time-consuming and costly due to the technical setups available today, therefore the examination may not be offered regularly. The purpose of this study was to evaluate the clinical performance of a pocket-size device for detecting DAN by measuring heart rate variability (HRV). Ten healthy young males and eight type 1 diabetes patients suffering symptomatic DAN were selected. The standardized spectral analysis equipment VariaPulse TF3 (Sima Media, Olumouc, Czechoslovakia) was used as a reference method for evaluating a prototype of the pocket-size device according to a specified protocol. HRV, inhalation/exhalation ratio (E:I) (deep breathing test), and 30:15 ratio (response going from lying to standing) were measured using both methods. Statistical calculations were performed. The correlation between the two devices was R(2) = 0.98 and R(2) = 0.81 when 30:15 ratio and E:I were measured, respectively. Bland-Altman plots showed suitable agreement between the two devices, substantiated by 95% limits of agreement of the differences of +/-0.014 and +/-0.033 when 30:15 ratio and E:I were measured, respectively. The pocket-size device was fully interchangeable with the hitherto-used, research-based setup. It proved highly suitable for ambulatory testing of autonomic nervous function and may facilitate screening for DAN according to Danish and international recommendations.