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Background: Few studies have evaluated the relationship between condom use and herpes simplex virus type 2 (HSV-2) and HSV type 1 (HSV-1) acquisition., Objective: To assess the relationship between condom use and acquisition of HSV-2 and HSV-1 among men and women., Design: Analysis of data collected as part of a clinical trial of an ineffective candidate vaccine for HSV-2., Setting: Sexually transmitted disease clinics., Participants: Men and women at risk for HSV-2 acquisition, defined as having 4 or more sexual partners or having a sexually transmitted disease in the past year., Measurement: Acquisition of HSV-2 and HSV-1 as measured by viral culture or change to positive HSV serostatus., Results: Of 1843 participants, 118 (6.4%) became infected with HSV-2. In multivariate analyses, participants reporting more frequent use of condoms were at lower risk for acquiring HSV-2 than participants who used condoms less frequently (hazard ratio, 0.74 [95% CI, 0.59 to 0.95]); categories of increasing condom use were 0% to 25%, 25% to 75%, and greater than 75% of sexual acts. Nineteen (2.9%) of 659 participants at risk for infection with HSV-1 became infected. No statistically significant association between condom use and infection with HSV-1 was found (hazard ratio, 0.79 [CI, 0.48 to 1.31])., Limitations: Use of condoms was measured by self-report, and persons who used condoms may have differed from those who did not., Conclusions: Consistent use of condoms is associated with lower rates of infection with HSV-2 and should be routinely recommended.