2.2. Preliminary confirmation and virus isolation
DNA was extracted from scab suspensions using QIAGEN Blood and tissue
DNA isolation kit. Isolated DNA was subjected to CaPV-specific PCR as
described earlier (Ireland and Binepal, 1998). After preliminary
confirmation, Vero cells grown in EMEM supplemented with antibiotics and
10% fetal calf serum were infected with filtered scab suspension using
the adsorption method. The cells were observed daily for the appearance
of cytopathic effect (CPE). After preliminary isolation of LSDV into
Vero cells, further propagation was continued up to seventy passages
(P70).