Coleus scutellarioides (L.) Benth., also known as coleus, is a globally important ornamental plant. The most recently discovered viral pathogen of coleus is Coleus vein necrosis virus (CVNV). It was first detected in North America in 2007, and subsequently in Croatia (Europe) in the Botanical garden of the Faculty of Science, Zagreb in 2018. In September 2020, two new coleus plants in the Botanical garden were observed to have virus-like symptoms. The aim of this work was to confirm CVNV as an agent infecting these two coleus plants and compare them to previous isolates from the Botanical garden. The CVNV infection was confirmed by biotest on experimental plants and by reverse transcription and polymerase chain reaction (RT-PCR) amplification and sequence analysis of a region of the viral coat protein gene. One of the isolates was examined by transmission electron microscopy and possible cytopathic effects were identified. Both new CVNV isolates failed to infect all of the known host species, but they didn’t differ from the previous isolates from Zagreb in other biological characteristics (symptomatology, growth conditions in planta). Phylogenetically, one of the new isolates was quite similar to older ones, while the other differed substantially. Further research is needed to see if the differences have biological or methodological basis.