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(1) Background: Detection of asymptomatic or subclinical human coronavirus SARS-CoV-2 infection in healthcare workers (HCWs) is crucial for understanding the overall prevalence of the new coronavirus and its infection potential in public (non-infectious) healthcare units with emergency wards. (2) Methods: We evaluated the host serologic responses, measured with semi-quantitative ELISA tests (IgA, IgG, IgM abs) in sera of 90 individuals in Hospital no. 4 in Bytom, 84 HCWs in the University Hospital in Opole and 25 in a Miasteczko Śląskie local surgery. All volunteers had negative RT-PCR test results or had not had the RT-PCR test performed within 30 days before sampling. The ELISA test was made at two different time points (July/August 2020) with a 2-weeks gap between blood collections to avoid the "serological window" period. (3) Results: The IgG seropositivity of asymptomatic HCWs varied between 1.2% to 10% (Opole vs. Bytom, p < 0.05; all without any symptoms). IgA seropositivity in HCWs was 8.8% in Opole and 7.14% in Bytom. IgM positive levels in HCWs in Opole and Bytom was 1.11% vs. 2.38%, respectively. Individuals with IgA and IgM seropositivity results were observed only in Opole (1.19%). More studies are needed to determine whether these results are generalizable to other populations and geographic as well as socio-demographic locations. (4) Conclusions: 100% of IgG(+) volunteers were free from any symptoms of infection in the 30 days before first or second blood collection and they had no awareness of SARS-CoV-2 infection. Asymptomatic HCWs could spread SARS-CoV-2 infection to other employees and patients. Only regular HCWs RT-PCR testing can reduce the risk of SARS-CoV-2 spreading in a hospital environment. The benefit of combining the detection of specific IgA with that of combined specific IgM/IgG is still uncertain.