A reduction in vascular access flow poses a risk for thrombosis. We present a new technique to measure vascular access flow during dialysis based on extracorporeal temperature gradients, and their changes, on reversing the extracorporeal bloodlines without having to inject an indicator. Fistula temperatures were measured by the blood temperature monitor with normal line position and after manual switching of the bloodlines using the same extracorporeal blood flow. The access flow by our temperature gradient method (TGM) was compared to access flow derived by saline dilution with measurements in the same patients repeated in subsequent weeks. In 70 pairs of TGM and saline dilution measurements in 35 patients, the repeatability of the TGM measurements was not significantly different from that of saline dilution. There was a highly significant correlation between the two techniques with an acceptable confidence level for limits of agreement for the difference between them. It took about 9 min to complete the TGM method and about 5 min for saline dilution. Our studies show that the novel TGM method showed excellent agreement and reproducibility with the saline dilution method without the need for indicator dilution.