Study on improving the plaque forming test for teaching purpose
Objective This experiment aimed to improve the"plaque test"in teaching by replacing the infecting virus to obtain more distinct"plaque test"results,so that it could better serve undergraduate experimental teaching.Methods Infect Vero cell in 12 well cell culture plate was infected with a serial concentration of Zika virus and dengue virus for 2 h in 37 ℃,and then 1％MC Eagle medium was used for the conducting cover step for 6-7 days at 37 ℃.After incubation,the infected cells were fixed and stained;cells were then washed with flowing water before observing the results.Results After staining,round,clear and complete plaques with neat edges could be seen and counted by bare eyes under the very bright violet background.The non-dead and exfoliated Vero cells infected with Zika virus were stained with crystal violet,and the purple background was obvious.Colorless plaques produced by Zika virus-infected cells could be seen with the naked eye.The plaques were round,clear and complete,with neat edges.The number of plaques increased with the increase of virus concentration,and the trend of the number of plaques produced in multiple wells was consistent.Unshedded Vero cells infected with dengue virus were stained purple,forming a brighter purple background.Dengue virus-infected cells produced colorless plaques,but the number of plaques formed was small,and were not consistent in duplicate wells.As the virus dilution factor increased,the number of plaques produced had no obvious decreasing trend.Conclusion Infected with Zika virus,a more representative plaque forming test result could be achieved which was more applicable for teaching purpose.
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