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The objective of this study was to determine if circumcision in early childhood affects the risk of acquiring herpes simplex virus type 2 (HSV-2) infection.Study members were born in 1972–1973 in Dunedin, New Zealand. Circumcision status was sought at age 3, when the cohort was established. Information about sexual behavior was obtained at ages 21 and 26. Serum was tested for HSV-2 antibodies at age 26 for 435 men (82.9% of the surviving cohort).Of eligible men, 40.2% had been circumcised. The prevalence of HSV-2 antibodies was 7.3% in uncircumcised men and 7.4% in circumcised men. Social and sexual factors were very similar between the 2 groups and adjustment had no effect on the association (odds ratio, 1.1; 95% confidence interval, 0.46–2.5). Seroconversion rates according to years since first sexual intercourse were 0.85 and 0.86 per 100 person-years for uncircumcised and circumcised men.The results support a lack of association between circumcision status and HSV-2 acquisition, although a small effect cannot be ruled out.