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Background Small private online course (SPOC) combined with the flipped classroomis established in higher education as an innovative tool for improvingstudents’ learning efficiency.To investigate the impact of the SPOC-based flipped classroomapproach, we implemented this reform pedagogy on anbasic immunology course for medical technology students. Methods The undergraduate medical technology students of Mahidol Universityenrolled SPOC platform for self-active online learning in the detection of immunological reactions and applications and then attended the face-to-face teaching activity in the classroom. The learning improvement was evaluated by conceptual test (CT)throughout this reform curriculum. Furthermore, students were required to fill in a questionnaire to reflect their perceptions and motivations. Results A significant increase in the overall score data of the mid- and the post-conceptual test was found. The result indicated that SPOC based flipped classroom method improved university students’learning knowledge in the detection of immunology reactions and applications. From the perception and motivation questionnaires, students had a positive attitude in which they had more practical and meaningful activities with this reform course. Students revealed that better interest and enjoyment with less anxiety could induce them to connect the topic to their career in the future. Conclusions TheSPOC-based flipped classroom approach effectively encouraged students’learning improvement and positively impacted their learning perceptions and motivations on fundamental immunology study in the field of medical technology education.