Authors : K. M. D. Islam , T. Ahmed , M. M. Ahasan , M. M. Billah , M. E. Islam and K. M. Hossain
Abstract: In the present experiment, adaptation of avian reovirus (Uchida, TS-17 AVREOV-1) on vero cell line and characterization of adapted virus by serological tests were carried out. For adaptation, the anchorage-dependent vero cell was first sub-cultured in Eagle`s minimum essential medium (EMEM) to form semi-confluent monolayer. This monolayer was then infected by the wild strain of reovirus. Subsequent passages of reovirus on vero cell line were performed by serially infecting the cell line with the virus. The first infection was done by the wild virus. The passage 1 (P1) virus was harvested and used for the next passage In this way, the viruses were given three serial passages on vero cell line where characteristic cytopathic effects (CPEs) were observed. During the third passage, the virus produced characteristic CPE for reovirus as in the adapted cell line. Syncytia and giant cells were clearly observed after 30-36 h, which were followed by plaque formation after 72 h of infection. The tissue culture infective dose 50 (TCID50) was performed in adapted vero cells to determine the infectivity titre of P3 adapted virus and it was found 10-5.5 TCID50, which indicated 10-5.5 times diluted vero cells adapted P3 virus were able to produce CPE in 50% of the cell cultures inoculated. The P1 virus showed less infectivity but highly virulent whereas the P3 virus became progressively more cytopathogenic or infective to vero cells. The purity of P3 adapted reovirus was detected by serum neutralization test (SNT), using specific anti-reovirus serum from rabbit. The homologous SNT against reovirus was 0.8x104 units per ml, whereas the non-homologous SNT was almost negative.
K. M. D. Islam , T. Ahmed , M. M. Ahasan , M. M. Billah , M. E. Islam and K. M. Hossain , 2003. Adaptation of Reovirus on Vero Cell Line . Journal of Animal and Veterinary Advances, 2: 425-429.