The object of the proposed paper is to design and analyze the performance of a non-contact heart rate variability (HRV) measuring device based on ultrasound transducers. The rationale behind non-contact HRV measurement is the goal of obtaining a means of long term monitoring of a patient’s heart performance. Due to its complexity as a non-contact measuring device, influential physical quantities, error source and other perturbations were thoroughly investigated. For medical purposes it is of utmost importance to define the target uncertainty of a measuring method from the side of physicians, while it is the role of scientists to realistically evaluate all uncertainty contributions. Within this paper we present a novelty method of non-contact HRV measurement based on ultrasound transducers operating at two frequencies simultaneously. We report laboratory results and clinical evaluations are given for healthy subjects as well as patients with known heart conditions. Furthermore, laboratory tests were conducted on subjects during a relaxation period, and after 1 min physical activity.