Background Opisthorchiasis caused by Opisthorchis viverrini is a serious health issue in the Mekong basin region, resulting in a high prevalence of cholangiocarcinoma. Nakhon Phanom province had the highest prevalence of O. viverrini infection in Thailand at 60% of the surveyed population in 2009, despite the attempted control of opisthorchiasis for >50 years. Knowing risk factors for O. viverrini infection in Nakhon Phanom may lead to improved control and prevention of opisthorchiasis. Objectives To determine risk factors for O. viverrini infection in Nakhon Phanom. Methods We conducted a cross-sectional survey in Nakhon Phanom province from February to March 2014. The community was selected using a stratified random sampling method, and then, participants were selected by systematic random sampling. Individuals ≥15 years old were included. Knowledge of O. viverrini infection, and attitudes and practice to avoid it were assessed using a questionnaire. O. viverrini infection was determined by stool examination with a formalin–ether concentration method. Factors associated with the infection were determined using multivariate logistic regression analysis. Results Of the 134 participants, 75 (56%) were infected with O. viverrini. In the multivariate logistic regression analysis, 3 independent factors were associated with O. viverrini infection: age ≥ 55 years, odds ratio (OR) adjusted 6.36 (95% confidence interval (CI) 1.28–31.66); consumption of chopped raw-fish salad (koi pla), OR adjusted 28.74 (95% CI 3.59–230.24); and perceived susceptibility, OR adjusted 0.15 (95% CI 0.03–0.74). Conclusions Age ≥ 55 years, consuming koi pla, and perceived susceptibility were independently associated with O. viverrini infection in Nakhon Phanom.