This study used a JDI 5-factor scale (Job Descriptive Index), reliability analysis method based on Cronbach’s Alpha coefficient, exploratory factor analysis (EFFA), linear regression analysis, T-test and one-way analysis of variance (One-way ANOVA) to achieve the research objectives. The research’s finding indicates that 5 factors affecting the job satisfaction of tour guides is the nature of work, welfare and salary, superior support, training and promotion and work environment. In addition, the findings of the research determine that the factor that has the strongest impact on job satisfaction of tour guide is the factor of superior support. The research also recommends solutions to increase the job satisfaction of tour guides in the Mekong Delta region based on the research findings.