Assay to Determine the Virus Infectious Dose of Drosophila C Virus in Cultured Insect Cells
Published: October 31, 2023
Abstract
Source: Yang, S. et al., Establishment of Viral Infection and Analysis of Host-Virus Interaction in Drosophila Melanogaster. J. Vis. Exp. (2019)
In this video, we demonstrate the cytopathic effect assay to determine the tissue culture infective viral dose of Drosophila C virus on Drosophila S2* cells. A range of viral concentrations are used to infect the cultured cells. The proportion of cells exhibiting structural changes is determined, and this data is used to calculate the tissue culture infective dose.
Protocol
1. Drosophila C virus load measure by CPE assay Pick 5 random virus-containing tubes to determine the average virus tissue culture infective dose (TCID50) by a cytopathic effect (CPE) assay (1 unit of TCID50= 0.7 plaque-forming unit [PFU]). NOTE: CPE refers to structural changes in host cells caused by viral invasion. The infecting virus causes lysis of the host cell or when the cell dies without lysis due to an inability to reproduce. O…
Declarações
The authors have nothing to disclose.
Materials
| 1.5 ml Microcentrifuge tubes | Brand | 352070 | |
| 10 cm cell culture dish | Sigma | CLS430167 | Cell culture |
| Cell Incubator | Sanyo | MIR-553 | |
| Centrifuge | Eppendorf | 5810R | |
| Centrifuge | Eppendorf | 5424R | |
| Drosophila Incubator | Percival | I-41NL | Rearing Drosophila |
| FBS | Invitrogen | 12657-029 | Cell culture |
| Schneider's Insect Medium | Sigma | S9895 | Cell culture |
| Statistical software | GraphPad Prism 7 | ||
| Flat bottom 96-well-plate | Sigma | CLS3922 | Cell culture |
| Cell Incubator | Sanyo | MIR-553 | |
| Microscope | Olympus | CKX41 |
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Assay to Determine the Virus Infectious Dose of Drosophila C Virus in Cultured Insect Cells. J. Vis. Exp. (Pending Publication), e21652, doi: (2023).