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The aim of this study was to investigate the seroprevalence of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in representative samples of the total former East German (FEG) and total former West German (FWG) populations. Type-specific testing with the HSV-1 (gG1) and HSV-2 (gG2) indirect ELISA was performed on an age- and sex-stratified random subsample of 3,792 sera collected during the 1997-1998 German National Health Survey. Weighted seroprevalence estimates were calculated for the total FEG and FWG populations. The overall age-standardised seroprevalence of HSV-1 was 82.6% in Germany, 85.5% (95%CI, 83.4-87.3%) in FEG and 81.8% (95%CI, 79.9-83.6%) in FWG. The overall seroprevalence of HSV-2 was 13.3% (95%CI, 11.8-14.9) in Germany, 16.5% (95%CI, 14.1-18.9) in FEG and 12.6% (95%CI, 10.7-14.5%) in FWG. The difference between FEG and FWG was largely due to the significantly higher age-adjusted seroprevalence of HSV-1 and HSV-2 in women (but not men) in FEG as compared to FWG. The HSV seroprevalence estimates in this study are consistent with results of previous less representative seroprevalence studies in Germany. Differences in HSV-2 seroprevalence between FEG and FWG suggest differences in sexual behaviour that warrant further investigation.