BackgroundTelemedicine system enables doctors and patients to communicate while staying afar which can be helpful for areas with lesser health facilities and at times of natural or health disasters. In developing countries like Bangladesh, telemedicine service offers the potential for wider health access and equity if effectively implemented. Therefore, the response and acceptance of care receivers who are the main beneficiaries of the service should be explored. As Dhaka University Telemedicine Programme (DUTP) is a non-profit University project-induced successful telemedicine service in Bangladesh, our study was conducted on the DUTP hotline-based telemedicine programme aimed to explore patients’ prior knowledge and response (experience, satisfaction and acceptance) about the service. MethodThe cross-sectional study was conducted by interviewing 200 participants over the phone with a structured questionnaire to analyze their knowledge and response. Participants were selected by randomization from the patient pool of hotline-based DUTP telemedicine service. The data was analyzed using SPSSv20. Results and discussionAmong the participants, 41% of total participants knew about telemedicine services before COVID-19. Average patient satisfaction was well above moderate level (p-value< 0.01; mean 3.88). 16.5% respondents mentioned about having any problem while availing the service where ‘treatment or service related problem’ was the most common. Significant association was found between people's ‘occupation’ and ‘knowledge before COVID-19’ (p = 0.002) indicating to the probable role of profession or occupation in molding people's health-service related knowledge. Patient-doctor communication [‘perception about doctor's adequate evaluation (Q3)’ and ‘understanding doctor's advice properly (Q4)’] was found to be significantly associated with ‘age’ and ‘location (division)’ while most respondents (around 90%) perceived the communication as effective. ‘Age’ had also an association with ‘treatment or service related problem’. 80.5% were willing to take the service in the future even by paying fees. All participants appreciated telemedicine service in general when they were asked about its possible inclusion in mainstream primary healthcare. ConclusionThe overall response of patients toward DUTP hotline-based telemedicine, in general, came out to be positive. Concerned authorities and policymakers may exploit this accepting attitude of people toward developing effective telemedicine services in order to ensure wider health and well-being of population.