Background: There is no published literature regarding the use of the digital stethoscope (DS) and computerized breath sound analysis in neonates, despite neonates experiencing a high burden of respiratory disease. We aimed to determine if the DS could be used to study breath sounds of term and preterm neonates without respiratory disease, and detect a difference in acoustic characteristics between them. Methods: A commercially available DS was used to record breath sounds of term and preterm neonates not receiving respiratory support between 24 and 48 hours after birth. Recordings were extracted, filtered, and computer analysis performed to obtain power spectra and mel frequency cepstral coefficient (MFCC) profiles. Results: Recordings from 26 term and 26 preterm infants were obtained. The preterm cohort had an average gestational age (median and interquartile range) of 32 (31-33) weeks and term 39 (38-39) weeks. Birth weight (mean and SD) was 1767 (411) g for the preterm and 3456 (442) g for the term cohort. Power spectra demonstrated the greatest power in the low-frequency range of 100 to 250 Hz for both groups. There were significant differences (P <.05) in the average power at low (100-250 Hz), medium (250-500 Hz), high (500-1000 Hz), and very high (1000-2000 Hz) frequency bands. MFCC profiles also demonstrated significant differences between groups (P <.05). Conclusion: It is feasible to use DS technology to analyze breath sounds in neonates. DS was able to determine significant differences between the acoustic characteristics of term and preterm infants breathing in room air. Further investigation of DS technology for neonatal breath sounds is warranted.