Tissues were harvested from infected mice into 500 μl PBS with approximately 20 glass beads in 2 ml Starstedt screw cap tubes. Tissues were bead beat in three 45 second cycles. Blood was collected and serum was collected from the clotted blood sample. A 20 μl sample of tissue lysates or sera were serially diluted in DMEM containing 5% FBS and PSG and added to 48-well plates containing a confluent monolayer of Vero cells and rocked at 37°C for 2 h. CMC in DMEM containing 5% FBS and PSG (250 μl per well) was overlaid and plates were incubated for 2 days at 37°C and then the cells were fixed with 3.7% formalin diluted in PBS and stained with 0.2% methyl blue dye for 15 minutes.

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