The COVID-19 pandemic has accelerated implementation of telemedicine in healthcare facilities for delivery of care. Healthcare providers’ acceptance of the telemedicine services is important for successful implementation of this new system. A questionnaire based on the Technology Acceptance Model (TAM) has been used to measure user acceptance of telemedicine service. The aim of this study was to translate and validate the English version of the questionnaire into Malay, to extend the availability and utilization of this questionnaire in Malaysia. A forward and backward translation of the questionnaire was conducted to produce the TAM in the Malay version (Malay-TAM). Panel experts assessed content validity. Internal consistency reliability was determined using Cronbach’s alpha. Confirmatory factor analysis based on structural equation modelling was performed to validate the factor structure. The questionnaire was then tested on and completed by 149 healthcare workers from several public health clinics across Malaysia. The Malay-TAM demonstrated good reliability with Cronbach’s alphas ranging from 0.823 to 0.912. Factor analysis showed good convergent validity but relatively poor discriminant validity. All five constructs were retained to preserve content validity. The findings suggest that the Malay-TAM can serve as a reliable and valid instrument to measure acceptance to telemedicine.