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Virus and cell culture

SL Salima S. Lalani
MA Mohd Ishtiaq Anasir
CP Chit Laa Poh
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Enterovirus (EV)-A71 strain 41 (5865/SIN/000009) was propagated in rhabdomyosarcoma (RD, ATCC# CCL-136) cells with Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with fetal bovine serum (FBS) and 1% penicillin-streptomycin antibiotics (PSA). Briefly, a monolayer of confluent RD cells in a T-75 flask was infected with EV-A71 virus and adsorption was allowed for 1 h at 37 °C in an incubator supplemented with 5% CO2. After incubation, the inoculum was removed and maintenance media (DMEM supplemented with 2% FBS) was added. Cytopathic effect (CPE) of RD cells was monitored over a period of 24–48 h and the virus was harvested by three freeze-thaw cycles. The virus was aliquoted and stored at -80 °C until further use. Vero (ATCC® CCL-81) cells were grown in DMEM supplemented with 10% FBS and 1% PSA.

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