Objective To accurately evaluate the influencing factors and urban-rural differences in the well-being among the elderly in Guizhou Province, so as to provide scientific basis and practical strategies for enhancing the well-being of the elderly. Methods Using stratified and convenience sampling methods, 1 500 elderly aged 60 and above who met the survey requirements were selected from six prefectures in Guizhou Province. The well-being of Older People Scale (WOOP) was applied for questionnaire surveys. Combined with the random forest model and the ordered logistic regression analysis, the importance evaluation and the urban-rural differences of the influencing factors on the well-being of the elderly in Guizhou Province were analyzed. Results The overall well-being level of the elderly in Guizhou Province was relatively high, and that in urban areas was slightly higher than that in rural areas. The mean value of subjective well-being among urban elderly was 4.21±0.65, while that of rural elderly was 4.11±0.75. The results of the random forest model and the ordered logistic regression analysis showed that physical health, mental health, social life, receiving support, acceptance and resilience, feeling useful, independence, and making ends meet were important influencing factors for the well-being of the elderly. The regression coefficient values of these factors all showed significance at the 0.001 level. Among them, the making ends meet (with a weight value of 0.231) was the core driving factor for the well-being of rural elderly. Meanwhile, mental health (with a weight value of 0.136) and physical health (with a weight value of 0.122) were the core driving factors for the well-being of urban elderly.Living situation (β=0.880, z=6.751, P<0.001) had a significant impact on the well-being of urban elderly but has no significant impact on that of rural elderly. Conclusion There are significant urban-rural differences in the influencing factors of the well-being of elderly in Guizhou Province. To enhance the well-being of elderly, it is necessary to start from multiple dimensions and implement precise policies. In rural areas, priority should be given to increasing the income of rural elderly. While in urban areas, efforts should be focused on optimizing the living situation of the elderly and building a comprehensive and multi-level medical and elderly care service support system.