Objective: To determine the relevant risk factors for the pathogenesis of periampullary carcinoma, so as to provide a theoretical reference for effective prevention and control of this conditioin. Methods: Using a 1:1 case-control matched design, 122 patients with periampullary carcinoma (case group) admitted during 2003 to 2014 and 122 patients without tumor or digestive disease admitted during the same period in Hunan Provincial People’s Hospital were enrolled. The possible risk factors were analyzed by conditional Logistic regression model. Results: Smoking, alcohol consumption and hepatitis B virus (HBV) infection were found to be the risk factors for periampullary carcinoma after controlling for confounding factors. The results showed that the risk of periampullary carcinoma increased in proportion with the increase in the number of cigarettes smoked per day, and odds ratio (OR) was 0.450 (95% CI=0.205–0.988), 0.500 (95% CI=0.092–2.730), 2.571 (95% CI=1.074–6.156) and 3.000 (95% CI=0.312–28.841) for those who did not currently smoke, smoked <20 cigarettes/d, 20–39 cigarettes/d and >40 cigarettes/d, respectively; the OR was 3.000 (95% CI=0.312–28.841), 65.289 (95% CI=0.006–70.239) and 4.500 (95% CI=0.972–20.827) for those who with alcohol consumption less than 40 g/d, 40-99 g/d and >100 g/d, respectively; the risk of periampullary carcinoma in cases with HBV infection was 3.25-fold higher than in those without history of HBV infection (95% CI=1.060–9.967). Conclusion: Heavy smoking, alcohol consumption and HBV infection are associated with increased risk of periampullary carcinoma.