Abstract A 1-year study involving 157 gastroenteritis samples was conducted to investigate the role of human astrovirus (HAstV) as a cause of gastroenteritis in Italian children aged < 2 years. The overall incidence of HAstV was 3.1%. Most cases occurred between March and May, and four of the five isolates were of the HAstV-1 type, the other being HAstV-3. Analysis of genetic variability showed that the three HAstV-1 isolates collected in 2000 clustered together, but separately from the 1999 isolate. The results indicated that HAstV should be considered as a potential diarrhoeal pathogen in Italian children.