Abstract: Respiratory Syncytial Virus (RSV) is a very important viral pathogen in children, immunocompromised and cardiopulmonary diseased patients and the elderly. Most of the published research with RSV was performed on RSV Long and RSV A2, isolated in 1956 and 1961, yet recent RSV isolates dier from these prototype strains. Additionally, these viruses have been serially passaged in cell culture, which may result in adaptations that aect virus–host interactions. We have isolated RSV from mucosal secretions of 12 patients in the winters 2016–2017 and 2017–2018, of which eight RSV-A subtypes and four RSV-B subtypes. Passage 3 of the isolates was assessed for viral replication kinetics and infectious virus production in HEp-2, A549 and BEAS-2B cells, thermal stability at 37 C, 32 C and 4 C, syncytia formation, neutralization by palivizumab and mucin mRNA expression in infected A549 cells. We observed that viruses isolated in one RSV season show dierences on the tested assays. Furthermore, comparison with RSV A2 and RSV B1 reveals for some RSV isolates dierences in viral replication kinetics, thermal stability and fusion capacity. Major dierences are, however, not observed and dierences between the recent isolates and reference strains is, overall, similar to the observed variation in between the recent isolates. One clinical isolate (BE/ANT-A11/17) replicated very eciently in all cell lines, and remarkably, even better than RSV A2 in the HEp-2 cell line.