This research is a qualitative study of the spiritual needs and practice of chronically ill children and their families in the Isan region of Thailand. The purposive sampling was 17 chronically ill children aged 10–18 years old and primary caregivers. Three instruments were used: a questionnaire, in-depth interview with questions adapted from the Spiritual Assessment Scale (SAS) by O'Brien, and non-participant observation. Field note-taking and voice recordings were used for data recording. Triangulation was used before analysis to test for credibility. Content analysis and thematic analysis techniques were used for data analysis. The five specific themes were stated by both children and caregivers. In addition, the spiritual needs and practice identified in this study were strongly related to Buddhism. All children and caregivers respected and believed in Buddha, and so Buddhism and Thai Isan traditions were crucial to their spiritual needs and were reflected in their religious practice.