In 2009, a sentinel surveillance system for Lyme disease (LD) was established in primary care in mainland France. Over the 2009–2016 period, 819 cases were reported by participating physicians. The estimated annual incidence of LD cases seen in general practice remained stable from 2009 to 2015, ranging from 41 cases (95% confidence interval, 95% CI: [31;51]) per 100,000 inhabitants in 2011 and 55 cases [43;67] per 100,000 inhabitants in 2013. It showed a significant increase in 2016 with 84 [70;98] reported cases per 100,000 inhabitants. Significant geographical heterogeneity was observed, with higher incidence rates in Limousin, Alsace, and Rhône-Alpes. The age distribution of cases showed a peak between 60 and 70 years of age. Among the cases reported by Sentinelle physicians, 95% presented with erythema migrans. It will be important to monitor whether this higher incidence in 2016 is confirmed in subsequent years.