Objective Near-infrared spectroscopy (NIRS) allows assessment of regional tissue oxygen delivery and extraction. There are doubts regarding reliability of gut NIRS measurements. This study assesses reliability of NIRS for monitoring gut oxygenation. Study design Splanchnic tissue haemoglobin index (sTHI), tissue oxygenation index (sTOI) and fractional tissue oxygen extraction (sFTOE) changes during blood transfusion were measured using NIRS and compared to stable control infants. Infants were grouped into 3 chronological age groups: 1–7, 8–28 and ≥29 days of life. Results sTHI, sTOI significantly increased, and sFTOE reduced following blood transfusion in all age group infants (n = 59), with no changes seen in control infants (n = 12). Baseline characteristics including gestational age and feed volumes did not differ between groups. Conclusion Gut perfusion measured by NIRS improved in infants who received blood transfusion, a change not seen in the control group, thus suggesting NIRS is a reliable method to measure splanchnic tissue oxygenation.