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ABSTRACT Background Revealing the clinical manifestations and associations of COVID‐19 before and after negative transition remains an area of significant uncertainty. The aim of this study is to investigate the clinical characteristics observed during and after Omicron infection among a specific population, namely healthcare workers (HCWs). Methods From November 4, 2022, to January 15, 2023, HCWs in our hospital were enrolled to document clinical symptoms, prevention, and treatment for COVID‐19 using a structured questionnaire. Results A total of 1101 HCWs were included, with SARS‐CoV‐2 infection detected in 78.20% (861/1101) during the observation period. The median duration for nucleic acid conversion was 8 days. Forty‐three symptoms were identified during SARS‐CoV‐2 infection (11 symptoms per individual). The common symptoms were fever, cough, headache, phlegm production, and fatigue (60.67%–83.29%). These symptoms can be further categorized into five groups: fever type, upper respiratory tract type, influenza type, digestive system type, and systemic type—all showing complex and diverse patterns. Following SARS‐CoV‐2 infection, a total of 19 symptoms were recorded including four newly emerged ones: reduced lung capacity, memory loss, lethargy, and inattention. Importantly, we observed a significant association between gastrointestinal symptoms during the nucleic acid positive phase and subsequent neuropsychiatric manifestations after negative conversion. Interestingly, females experiencing menstruation or pregnancy exhibited a higher risk of infection, while inhaled vaccines and thymosin demonstrated a protective effect against SARS‐CoV‐2 infection. Conclusions The clinical manifestations observed in HCWs during and after Omicron infection displayed intricate patterns, shedding new light on the complex interplay between SARS‐CoV‐2 and humans.