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…

Offenlegungen

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).

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