目的建立评价副溶血弧菌毒力的Raw-264细胞模型，并以此评价副溶血弧菌临床分离株RIMD2210633株与其重要致病相关基因opaR基因敲除株的毒力。方法以Raw-264细胞为靶细胞，RIMD2210633株及其opaR基因敲除株为效应细胞，通过优化条件，利用CytoTox96Non-RadioactiveCytotoxicityAssay建立细胞模型。结果以5倍于靶细胞的菌量感染靶细胞，37℃孵育1.5h，离心所得上清稀释10倍后，加底物液避光显色10min，反应条件最佳。opaR基因敲除后，菌株的Raw-264细胞毒性升高，并有统计学差异。结论利用副溶血弧菌标准强毒株RIMD2210633菌株建立了评价副溶血弧菌毒力的RAW-264细胞模型，并利用此平台比较了该毒株与其opaR基因敲除株的毒力，为研究副溶血弧菌的致病机制奠定了基础。

Objective To establish a Raw-264 cell model for evaluating virulence of Vibrio parahaemolyticus. Methods The CytoTox 96 Non-Radioactive Cytotoxicity Assay Kit was employed to test the Vibrio parahaemolyticus-induced cytotoxicity against Raw264 cells. Results The protocols were optimized as follows: Vibrio parahaemolyticus cells and Raw264 cells were incubated for 1.5 h at 37℃ with a MOI of 5; the mixture was then pelleted, and the 10-fold diluted supernatant was incubated with the substrate for 10 min. The cytotoxicity against Raw-264 cells determined for the opaR mutant was significantly higher than that for the wild-type strain. Conclusions A Raw-264 cellular model was established for evaluating virulence of Vibrio parahaemolyticus.