Background Human parvovirus B19 (B19V) is responsible of a wide clinical spectrum ranging from asymptomatic infection, through mild disease, up to life threatening one. Outbreaks are registered every three to four years and a recent international alert for a new outbreak has been released. Here is reported the experience of B19 virus circulation in a 600-bed tertiary care pediatric hospital in Rome, from 2018 to 2024. Methods This retrospective study involved a total of 9,695 blood samples (about 8,500 patients), 11 amniotic fluids (11 pregnant women) and 2,483 sera (about 2,000 patients), processed in the Virology Unit of Bambino Gesù Children’s Hospital in Rome for B19V direct and indirect detection. Results and Conclusions The 9.6% of blood samples investigated for B19V DNA resulted positive. The annual positivity rate ranged from 0.8% in 2023, to 9.8% in 2018 and 32.8% in 2024; the same epidemiology resulted from the analysis of the Immunoglobulins M and G anti-B19V. Focusing on the last year, 314 patients resulted positive for B19V DNA detection: 204/314 (65%) had a primary infection, 150/204 (73.5%) were hospitalized and 17/150 (11.3%) needed Intensive Care Unit (ICU) for cardiovascular, central nervous and gastrointestinal pathologies. Two patients died for myocarditis. Four amniotic fluids resulted positive. B19V typing of a subset of samples revealed the presence of only genotype 1A and a low intra-genotypic diversity between strains from severe and mild disease. In conclusion, in 2024 was observed a significant increase of B19V circulation with serious effects on clinical outcome and consequent hospitalization.