A serosurvey of 9,673 United States military personnel was conducted to estimate infection rates with Borrelia burgdorferi sensu stricto, which is the cause of Lyme disease in the United States. Initial screening of sera from 9,673 military personnel on active duty in 1997 was performed by enzyme-linked immunosorbent assay (ELISA); supplemental testing of all ELISA-positive sera was performed by Western blot. Initial screening identified 1,594 (16.5%) ELISA-positive samples, but only 12 (0.12%, 95% confidence interval [CI] = 0.05-0.19%) were confirmed by Western blot. Antecedent serum samples collected from 1988 to 1996 were available for 7,368 (76%) subjects, accounting for 34,020 person-years of observation. Just two of the nine Western blot-positive individuals for whom antecedent samples were available seroconverted during military service for an annual incidence rate of six seroconversions per 100,000 persons (95% CI = 0.7-21.5). The risk of Lyme disease in the U.S. military population was found to be low. Although there may be sub-groups of military personnel who could potentially benefit from vaccination, force-wide use of the Lyme disease vaccine is not warranted.