The use of the continuous cell line CaCo-2 as an in vivo amplification system for the detection of fastidious human enteric viruses is reported. CaCo-2 cells showed an increased sensitivity to laboratory strains of group A rotavirus 3, reovirus 3, astrovirus 1, poliovirus 1, coxsackievirus A 24, enterovirus 70, and adenovirus 5, 40 and 41, when compared to a routine host cell line for each virus. Nucleic acids from wild-type infectious rotavirus, astrovirus, and adenovirus 40 in stool samples of patients with acute gastroenteritis could be amplified after infection of CaCo-2 cells with trypsin-pre-treated virus inocula. Virus diagnosis was carried out subsequently by dot-blot hybridisation with specific cDNA probes. An amplification factor between 10 and 1,000x was obtained by infection of CaCo-2 cells, thus enabling specific detection of low numbers of a wide range of enteric viruses, and the differentiation between infectious and noninfectious particles.