Background The number of diagnostic assays for the detection of herpes simplex virus type 1 (HSV-1) antibodies has increased over the years. However, their performance characteristics could vary among global populations. Objective To investigate performance of two commercial ELISA kits, HerpeSelect® 1 ELISA and Euroimmun Anti-HSV-1 (gC1) ELISA (IgG); and two commercial immunoblot (IB)/Western blot (WB) assays, HerpeSelect® 1 and 2 Immunoblot IgG, and Euroimmun Anti-HSV-1/HSV-2 gG2 Euroline-WB (IgG/IgM); in detecting HSV-1 antibodies in a Middle East and North Africa (MENA) population. Study design Blood specimens were collected from blood donors in Doha, Qatar, June 2013–2016. Twenty specimens were randomly selected from 10 MENA nationalities (Egypt, Iran, Jordan, Lebanon, Pakistan, Palestine, Qatar, Sudan, Syria, and Yemen; total = 200), and tested for HSV-1 antibodies. Results Across all six comparisons between assays, positive percent agreement ranged between 95.7% (95% CI: 91.4–98.3%) and 100.0% (95% CI: 97.8–100.0%). Negative percent agreement ranged between 86.2% (95% CI: 68.3-96.1%) and 96.2% (95% CI: 80.4–99.9%). Overall percent agreement ranged between 95.7% (95% CI: 91.7–97.8%) and 99.4% (95% CI: 96.7–99.9%). Cohen’s kappa statistic ranged between 0.84 (95% CI: 0.73–0.95) and 0.98 (95% CI: 0.93–1.00). Compared against IB/WB, HerpeSelect® and Euroimmun had sensitivities and specificities >96% and >86%, respectively. Positive and negative predictive values were >97% and >83%, respectively. Conclusion The assays showed excellent concordance with one another, and with a high kappa statistic. The ELISA kits demonstrated robust diagnostic performance compared to the IB/WB assays. These findings support the assays’ utility in clinical diagnosis and research in MENA populations.