Background: The active knee extension (AKE) test commonly used to assess the flexibility of the hamstring muscles. Many researchers have tested the reliability of the AKE test; however, no published studies have examined the intrarater and interrater reliability of the AKE test using a PBU. Objects: The purpose of this study was to determine the intrarater and interrater reliability of the AKE test performed with a pressure biofeedback unit (PBU) on healthy subjects. Methods: Sixteen healthy male participants volunteered and gave informed consent to participate in this study. Two raters conducted AKE tests independently with a PBU. Each knee was measured twice, and the AKE testing was repeated one week after the first round of testing. Results: The interrater reliability’s intraclass correlation coefficients (ICC2,1) were .887∼.986 for the right knees and .915∼.988 for the left knees. In addition, the intrarater (test-retest) reliability (ICC3,1) values ranged between .820∼.915 and .820∼.884 for Raters 1 and 2, respectively. The values for the standard error of mesurement were low for all tests (.81∼2.97˚); the calculated minimum detectable change was 2.24∼8.21˚. Conclusion: These findings suggest that the AKE test performed with a PBU had excellent interrater and intrarater reliability for assessing hamstring flexibility in healthy young males.