Objectives: Hairstylists form an occupational group whose tasks involve repetitive and forced movements of hands and wrists, thus posing a risk of developing carpal tunnel syndrome (CTS). This study assessed the prevalence of and factors associated with CTS symptoms among hairstylists in Gaborone, Botswana. Material and Methods: A cross-sectional study was conducted using a self-administered questionnaire distributed among randomly selected hairstylists. The questionnaire gathered information on demographic characteristics, lifestyle, work-related characteristics and psychosocial factors. The Boston Carpal Tunnel Syndrome Questionnaire was used to determine the severity of CTS symptoms and its functional effects. Data were then analyzed using χ² and logistic regression models. The level of significance was determined at p < 0.05. Results: A total of 165 hairstylists took part in the study, with 92 (56%) of the respondents being females. The mean age (M±SD) of the respondents was 35.05±7.54 years with an age range of 22–63 years. Seventy-three (42.2%) hairstylists reported CTS symptoms, with the majority (73%) being females. Out of all the CTS cases, 53 (72.6%) and 16 (21.9%) had mild and moderate symptoms, respectively. Over 80% of the hairstylists did not know about CTS. Among individual factors, CTS symptoms were associated with being female (the odds ratio [OR] of 9.99, and the 95% confidence interval [CI]: 3.64–27.44), increasing age (OR 9.84, 95% CI: 2.74–35.36), the length of employment (OR 3.73, 95% CI: 1.39–9.95), hair washing (OR 2.88, 95% CI: 1.41–5.85), an awkward posture (OR 2.52, 95% CI: 1.03–6.19), and the use of a great muscular effort when performing a task (OR 2.39, 95% CI: 1.01–5.72). Perceived heavy workload and stressful work were also risk factors. Conclusions: The results suggest a high prevalence of CTS among female hairstylists in Gaborone, and also point out that individual, work-related and psychosocial factors are associated with this syndrome. Future large-scale research is needed to establish the extent of CTS countrywide to influence policy-making. Currently, CTS is not listed amongst occupational health diseases in Botswana. Int J Occup Med Environ Health. 2021;34(3):437–50