BackgroundRotavirus is considered worldwide as one of the most important viral gastrointestinal infections, resulting in potentially life-threatening diarrhoea and death in children under the age of 5 years. Rotavirus can survive and remain infectious for long periods outside of the human body and can be easily transmitted via environmental surfaces.MethodStool specimens that had been collected and stored since 2010/2011 at 2°C – 8°C instead of −20°C or −80°C were analysed to determine the viability of rotavirus in these specimens after 6 years of improper storage. The specimens were analysed using simple enzyme immunoassay (EIA) methods from two different suppliers at different times throughout the period (2012–2017).ResultsThe analysis showed similar detection results for the two EIA kits.ConclusionThe rotavirus can be detected after several years of incorrect storage with EIA kits.