Agricultural tourism is a type of tourism, which is highly concerned to develop in the Mekong Delta, Vietnam. At agricultural tourism destinations, homestay is a typical form of accommodation, an inseparable product of this tourism type. To increase profits for homestays at agricultural tourism destinations and tourists’ satisfaction, and also to develop sustainably, it is necessary to research and improve the service quality (SQ) at those homestays. Thus, the aim of this study is to assess the quality of homestay services at agricultural tourism destinations in the centre of the Mekong Delta, Vietnam. The SQ model, SERVQUAL combined with the IPA model to solve the research issue. Six criteria of the service quality including tangibles, reliability, responsiveness, assurance, empathy, and cultural performance were applied to evaluate in the present research. A structured questionnaire was used for data collection. From June to December 2023, a survey of 258 tourists was conducted at homestays belonged to agricultural tourism destinations in Can Tho (CT) City by physically and online. Descriptive statistics and paired-samples T-tests were used to analyze the collected data. The study concluded that all criteria were highly evaluated towards the significance, whereas the productivity when homestay owners conducted was found to be less-than-expected. Besides, international tourists showed higher evaluation towards the significance and the productivity of those homestays than that of domestic ones. The research result implied that homestay owners should plan to improve the SQ at their homestays in order to be consistent with the reality and suitable for the target tourist groups.