Background Healthcare workers are one of the occupational groups that experience neck pain most frequently. The aim of this study is to investigate the immediate effects of Post Isometric Relaxation (PIR) Technique on pain threshold, pain severity and joint range of motion of the cervical region in healthcare workers with neck pain. Methods A total of 30 volunteer male and female hospital workers aged 20-45 years, with complaints of neck pain for at least six months and with neck pain as the most prominent complaint, were randomly divided into two groups (Post Isometric Relaxation Technique and Control Group). Control group participants received classical massage to the cervical and upper thoracic region. PIR group participants received a single session of PIR Technique with 10 repetitions to the M.supraspinatus, the upper part of the M.trapeze, M. scaleni and the M.levator scapula muscles in addition to massage. Pain severity, pain threshold and range of motion were measured at baseline and immediately after the treatment programme. Results Comparisons were made using both independent samples test and Mann-Whitney U tests. In-group comparison, participants of both groups improved in all measurments immediately after the treatment programme (p < 0,005). There were significant differences in a favour of PIR Group in inter-group comparison across the board (p < 0,01). Conclusions The results of the study suggest that in addition to application of massage, PIR Techniques improve pain intensity, pain threshold and range of motion immediately after the programme in non-specific neck pain.