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Background: This study was conducted to evaluate the anti-SARS-CoV-2 IgM and IgG antibodies among healthcare workers in Guilan.Methods: This cross-sectional study was conducted on 503 healthcare workers. Between April and May 2020, blood samples were collected from the healthcare workers of Razi Hospital in Rasht, Guilan, Iran. Enzyme-linked immunosorbent assay (ELISA) was used for the detection and quantitation of anti-SARS-CoV-2 IgM/IgG antibodies by using kits made by Pishtaz Teb Company, Tehran, Iran.Results: From a total of 503 participants, the result of the anti-SARS-CoV-2 IgM antibody test was positive in 28 subjects (5.6%) and the anti-SARS-CoV-2 IgG antibody test was positive in171 subjects (34%). Participants in the age group of 35-54 years were significantly more likely to have a positive anti-SARS-CoV-2 antibody test than the age group of 20-34 years (odds ratio = 1.53, 95% CI: 1.04-2.25, P=0.029). Also, physicians were significantly more likely to have a positive antibody test than office workers (odds ratio = 1.92, 95% CI: 1.04-3.54, P=0.037). The wide range of symptoms was significantly associated with the positive anti-SARS-CoV-2 antibody test. The most significant association was observed between fever and a positive anti-SARS-CoV-2 antibody test (odds ratio = 3.03, 95% CI: 2.06-4.44, P < 0.001).Conclusion: The results of the current study indicated that the seroprevalence of COVID-19 was high among healthcare workers of Guilan Province. It seems that this finding was due to the earlier exposure to COVID-19 and the lack of awareness and preparedness to deal with the pandemic in Iran, compared to other countries. [ABSTRACT FROM AUTHOR]