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The study aims to investigate the prevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection among ophthalmology unit staff throughout the first and second waves of the outbreak, in order to verify the effectiveness of the measures adopted in containing the contagion. A retrospective observational study was conducted involving staff members, who received a naso/oropharyngeal swab when complaining of SARS-CoV-2 symptoms and once a month as a screening measure. They were tested for SARS-CoV-2 antibodies as a screening measure during the first and the second wave. Clinical activities performed during the outbreak were compared with those performed during the same period in 2019 and correlated with the number of coronavirus disease-2019 eye care workers. Analysis included 25 workers. Clinical infection was 0% and 12% whereas the prevalence of SARS-CoV-2 antibodies ranged from 4% to 8% in the first and second wave, respectively. The increase in the prevalence of SARS-CoV-2 infection between the first and the second wave was not significant (1/25 vs 3/25, P = .6092). Clinical activities significantly decreased during the first wave compared with the same period in 2019 (3256 vs 10,075, P