Canine distemper virus (CDV) is a common infectious agent that affects respiratory and nervous systems
in canine, pet and wildlife worldwide. The virus isolation using cell culture is one of laboratory diagnostic
methods for detection of virus infections. Many different types of cultured cells can be used for isolation of
distemper virus, which provides the different outcome. This study aimed to evaluate the efficacy of dual cellculture
system, Crandell-Rees feline kidney (CRFK) and HeLa cells for virus isolation. A total of 13 clinical
specimens were taken by nasal and oropharynx swab from dogs with clinical signs compatible with CDV
infection. These specimens were positively confirmed CDV by using Reverse transcriptase - polymerase chain
reaction (RT-PCR) for viral genome detection. The CDV RT-PCR positive specimens were then taken to isolate
in both cell-lines for five passages, and were confirmed by RT-PCR again. By using CRFK and HeLa cells, the
successful viral isolation with CDV positive specimens was recorded at 7/13 (53.84%) in CRFK and 6/13 (46.15%)
in HeLa cells, respectively. The dual cell-culture system showed an increase of the successful isolation at 8/13
(61.53%). Our results suggested that the application of dial cell-culture system was more effective than using
single cell-culture for isolation of canine distemper virus.