Passage 1 of Ebola virus H.sapiens-tc/GIN/2014/Makona-C05 virus (Rocky Mountain Laboratory, National Institutes of Health, Hamilton, MT, USA) was used to generate passage 2 virus stock in Vero E6 cells
Vero E6 cells
(ATCC[R] number CRL-1586[TM]) were maintained in Modified Eagle Medium (MEM) supplemented with 10% heat-inactivated fetal calf serum (FCS) at 37[degrees]C in a humidified atmosphere of 5% C[O.sub.2].
The mosquito suspension was centrifuged; Millipore filtered (pore size = 0.22 pm), diluted serially (10-fold) and titrated in Vero E6 cells
Vero E6 cells
(ATCC CCL81) were grown and maintained in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% and 2% fetal bovine serum (FBS), respectively.
In addition, infectious arenavirus was recovered from materials from the 14-year-old patient by cultivation of the virus in monolayer cultures of Vero E6 cells
; virus isolation attempts on materials from the 30-year-old patient are under way.
In Vero E6 cells
, which had been inoculated with the specimen, diffuse cytopathic effects typical for DENV-1 developed but at a much later time than for DENV-1-positive plasma specimens from other patients; this finding raised the possibility that DENV-1 had either mutated to reduced replication fitness or that the cells were co-infected with >2 incompatible viruses that were interfering with the replication of each other.
By using Vero E6 cells
(American Type Culture Collection), we isolated Zika virus from 9 samples (8 serum, 1 urine).
Virus stocks were grown in Vero E6 cells
and titrated by using a 50% tissue culture infectious dose ([TCID.sub.50]) assay (9).
These 2 viruses were propagated on Vero E6 cells
, titered on Vero E6 cells
, and stored in liquid nitrogen.
No direct antiviral effect for lamivudine was observed at concentrations [less than or equal to] 320 [micro]mol/L in Vero E6 cells
To isolate SFTSV, we inoculated subconfluent mono layers of Vero E6 cells
with the RT-PCR-positive semm.
Virus isolation was attempted in Vero E6 cells
. We tested serum samples for neutralizing antibody titers using a validated MERS-CoV spike pseudoparticle neutralization test (7) (online Technical Appendix).