I propagated CHIKV on C6/36 before and harvested on different days of infections, though the results of immunofluorescence assay are highly positive for infection, I cannot get good plaque results. The titres are so less that I couldn't do the replication kinetics. Now, I moved to propagate stock CHIKV in vero cell. How to get a high virus titre of CHIKV propagated on Vero cells?
Previously, I performed CHIKV propagation on Vero C1008 cells in DMEM 5% using T25 flasks. To get a high virus titer, I did several passaging of CHIKV in a T25 flask. During each passage, the time required to observe cytopathic effect (CPE) was reduced. Eg: on the 1st passage, CPE was observed at 72 hours after CHIKV inoculation so I collect the supernatant containing the virus, take 200 microliter of the supernatant and inoculate the virus in a new T25 flask containing 80% confluent Vero C1008 cells. During this 2nd passage, I noticed the CPE at ~60 hours post inoculation. Repeat this passaging step a few times and finally collect the supernatant when CPE reaches about 48 hours. Make sure to always observe the cells so as not to miss the CPE. At this point, I inoculate CHIKV into several T25 flasks to make a large stock of CHIKV. Hope this helps.
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