The Dynavision D2™ Mode A test (ModeA) is a 1-minute reaction time (RT) test commonly used in sports science research and clinical rehabilitation. However, there is limited data regarding the effect of repeated testing (ie,training) or subsequent periods of no testing (ie,detraining) on test-retest reliability and RT performance. Therefore, the purpose of this study was to examine the test-retest reliability, training, and detraining effects associated with the D2™ ModeA test. Repeated measures/reliability. Twenty-four recreationally active men and women completed 15 training sessions consisting of 2 ModeA tests per session (30 tests). The participants were then randomized to either 1 or 2 weeks of detraining prior to completing 15 retraining sessions (30 tests). The training and retraining periods were separated into 10 blocks for analysis (3 tests per block). The number of hits (hits) and the average RT per hit (AvgRT) within each block were used to determine RT performance. Intraclass correlation coefficients, SEM, and minimum difference were used to determine reliability. Repeated-measures analysis of variance/analysis of covariance were used to determine training and detraining effects, respectively. The ModeA variables demonstrated excellent test-retest reliability (intraclass correlation coefficient2,3 > .93). Significant improvements in hits and AvgRT were noted within training blocks 1 to 5 (P < .05). No further improvements in RT performance were noted between training blocks 6 through 10. There was no effect of detraining period on RT. The RT performance was not different between blocks during retraining. It appears that 15 tests are necessary to overcome the training effect and establish reliable baseline performance for the ModeA test. Detraining for 1 to 2 weeks did not impact RT performance. The authors recommend that investigators and clinicians utilize the average of 3 tests when assessing RT performance using the D2 ModeA test.