Introduction The most common work-related musculoskeletal disorder worldwide is neck pain, especially among individuals who extensively use computers in a seated position. One biomedical cause of this neck pain is the prolonged activity of the muscles around the neck. Electromyography (EMG) has been used to evaluate the frequency and intensity of muscle activity. "Relative rest time (RRT)" is an index derived indicating the proportion of time below a certain threshold to the total task time. This study aimed to investigate the measurement errors and minimum detectable change (MDC) of RRT of the pericervical muscles during prolonged typing tasks in individuals with neck symptoms and to examine the differences in measurement errors at 3 µV and 6 µV thresholds. Methods This test-retest reliability study was conducted twice with a one-week interval to examine measurement errors of the RRT using surface EMG. The number of participants was set to 30 subjects who had neck symptoms with a Neck Disability Index of 16% or higher. The primary outcome measure was RRT of the following: the pericervical muscles of the right side during a 60-minute typing task; the splenius capitis muscle, upper trapezius (UT) muscle, middle trapezius muscle, sternocleidomastoid muscle, serratus anterior muscle, longissimus muscle, and pectoralis major muscle. RRT was calculated as the percentage of time that muscle activity was below the threshold for more than 0.250 seconds continuously during a 60-minute typing task. The standard error of measurement (SEM) and MDC were calculated with the two thresholds of 3 µV and 6 µV. The SEMs of the two thresholds were compared using a paired method. Results Ultimately, the data of 26 participants were analyzed. The SEM (MDC) values of the RRT at the 3 µV and 6 µV thresholds were 19.22 (53.27) and 9.52 (26.39) for the splenius capitis muscle, 3.24 (8.97) and 0.38 (1.05) for the sternocleidomastoid muscle, 15.47 (42.88) and 18.79 (52.08) for the UT muscle, 21.28 (58.99) and 2.28 (6.32) for the middle trapezius muscle, 13.67 (37.90) and 11.64 (32.27) for the serratus anterior muscle, 16.81 (46.60) and 3.32 (9.20) for the longissimus muscle, and 8.97 (24.87) and 4.24 (11.74) for the pectoralis major muscle, respectively. The SEMs of the RRT with the 6 µV threshold were statistically significantly lower than those with the 3 µV threshold in all pericervical muscles, except for the UT muscle. Conclusion This study identified the SEM and MDC of the RRT for the pericervical muscles during prolonged typing tasks in individuals with neck symptoms. Except for the UT muscle, the SEMs of the RRT with the 6 µV threshold were statistically smaller than those with the 3 µV threshold. Therefore, when using the RRT in intervention studies that aim to reduce muscle activity during typing in those with neck symptoms, the 6 µV threshold measurement would be recommended for the RRT of the pericervical muscles except for the UT muscle.