Post-COVID-19 syndrome (PCS) is a common outcome of severe COVID-19 infection; however, less is known about PCS following mild COVID-19. Healthcare workers (HCWs) are more susceptible to acquiring COVID-19 and potentially suffering physical and psychological morbidity secondary to their role. We surveyed HCWs at four hospitals in northeast England at two timepoints during the pandemic, assessing physical and psychophysiological symptoms of PCS, alongside associated factors, whilst also testing for COVID-19 status by SARS-CoV-2 serology and reviewing evidence of infection from previous PCR nasopharyngeal swabs. Of the 379 participants at baseline and 250 HCWs recruited 18 months later, 46% and 64% (respectively) had evidence of previous COVID-19, with no significant associations between COVID-19 status or demographics and symptom scores or self-described PCS. Depression and fatigue were more common later in the pandemic. Furthermore, 20% self-described having PCS, 34% of whom had no evidence of previous COVID-19. Scores for fatigue, pain, mobility, anxiety, and depression were significantly worse in the PCS group compared to those without ongoing symptoms. Significant proportions of HCWs continued to suffer debilitating symptoms during the later pandemic period, although a significant minority had no evidence of previous COVID-19 infection, suggesting that other factors may be involved in their symptomatology.