1.Reverse genetic virus from Dr. 王貞仁 (成功大學); 2. Strain Taiwan/3446/02 containing double mutations on NP(K31R-S450G); 3. Other related strains are available from NIDB.
Seed cells on culture dishes 24-48h before viral infection and the infection should be conducted when cultures reach 70-90% confluency. After removing the cell culture medium, the inoculation is performed using a small volume (e.g. 1.5-3 ml per 10 cm dish) and an appropriate M.O.I. (e.g. 0.1). Follow 1-2 h of adsorption at a 35℃, humidified, 5 % CO2 incubator, rocking culture dish every 15-30 min to redistribute the inoculum. End adsorption by adding virus growth medium. Harvesting virus until CPE is progressed through 80%