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The purpose of this study was to determine the intra-rater, inter-examiner and inter-observer reliability of biceps femoris long head (BFlh) tendon strain using ultrasound imaging. Nineteen patients (age: 20.4 +/- 0.35 y) were tested twice with a 1-wk interval. Each session included passive stretching from three different hip positions. Tests were performed independently by two examiners while BFlh tendon displacement (mm) and strain (%) were manually extracted from ultrasound video footages by two observers. Intra-rater comparisons revealed an intra-class correlation coefficient (ICC2,1) range of 0.87 to 0.98 and a variability less than 4.74%. Interexaminer comparisons revealed an ICC2,1 range of 0.83 to 0.99 and less than 4.69% variability. Inter-observer ICCs ranged from 0.93 to 0.97 with variability less than 4.89%. Using a well-defined scanning protocol, two experienced examiners attained high levels of intra-rater agreement, with similarly excellent results for inter-rater and inter-observer reliability for BFlh tendon displacement and strain. (E-mail: ekellis@phed-sr.auth.gr) (C) 2017 World Federation for Ultrasound in Medicine & Biology.