Objective: To provide a practical guide to appropriate statistical analysis of a reliability study using real-time ultrasound for measuring muscle size as an example. Design: inter-rater and intra-rater (between-scans and between-days) reliability. Subjects: Ten normal subjects (five male) aged 22-58 years. Method: The cross-sectional area (CSA) of the anterior tibial muscle group was measured using real-time ultrasonography. Main outcome measures: Intraclass correlation coefficients (ICCs) and the 95% confidence interval (CI) for the ICCs, and Bland and Altman method for assessing agreement, which includes calculation of the mean difference between measures ((d) over bar), the 95% CI for (d) over bar, the standard deviation of the differences (SDdiff), the 95% limits of agreement and a reliability coefficient. Results: inter-rater reliability was high, ICC (3,1) was 0.92 with a 95% CI of 0.72 --> 0.98. There was reasonable agreement between measures on the Bland and Altman test, as (d) over bar was -0.63 cm(2), the 95% CI for (d) over bar was -1.4 --> 0.14 cm(2), the SDdiff was 1.08 cm(2), the 95% limits of agreement -2.73 --> 1.53 cm(2) and the reliability coefficient was 2.4. Between-scans repeatability was high, ICCs (1,1) were 0.94 and 0.93 with 95% Cfs of 0.8 --> 0.99 and 0.75 --> 0.98, for days 1 and 2 respectively. Measures showed good agreement on the Bland and Altman test: (d) over bar for day 1 was 0.15 cm(2) and for day 2 it was -0.32 cm(2), the 95% CIs for (d) over bar were -0.51 --> 0.81 cm(2) for day 1 and -0.98 --> 0.34 cm(2) for day 2; SDdiff was 0.93 cm(2) for both days, the 95% limits of agreement were -1.71 --> 2.01 cm(2) for day 1 and -2.18 --> 1.54 cm(2) for day 2; the reliability coefficient was 1.80 for day 1 and 1.88 for day 2. The between-days ICC (1,2) was 0.92 and the 95% CI 0.69 --> 0.98. The (d) over bar was -0.98 cm(2), the SDdiff was 1.25 cm(2) with 95% limits of agreement of -3.48 --> 1.52 cm(2) and the reliability coefficient 2.8. The 95% CI for (d) over bar (-1.88 --> -0.08 cm(2)) and the distribution graph showed a bias towards a larger measurement on day 2. Conclusions: The ICC and Bland and Altman tests are appropriate for analysis of reliability studies of similar design to that described, but neither test alone provides sufficient information and it is recommended that both are used.
