影响轮状病毒半数细胞感染剂量法滴定的因素分析

Factors affecting rotavirus quantization by cell culture infection dose-50

目的:确定影响轮状病毒感染增殖过程的因素,以提高滴度检测结果的重复性和可靠性。方法:采用半数细胞感染剂量法(CCID50)结合ELISA(CCID50-ELISA)判断病毒在细胞上有无复制来检测病毒滴度。结果:不同细胞代次、接种浓度、培养时间、培养液种类和接种细胞孔数均会对病毒滴定结果造成显著影响。结论:轮状病毒滴度检测的线性和准确性与细胞状态与培养环境相关。CCID50-ELISA法检测轮状病毒滴度有良好的重复性和耐用性。

Objective： To confirm the factors that affect replications of rotavirus and improve the repeatability and reliability of titration. Methods： The virus titers were determined using enzyme-linked immunosorbent assayformat cell culture infectious dose-50 test for titrating rotavirus （CCID50-ELISA）. Results： All the factors, inclu- ding passages of cells, MOI and incubation time of infected cells, types of culture media and the number of wells inoculated with viruses significantly have impact the results of virus titration. Conclusion： CCID50-ELISA has high repeatability and robustness for rotavirus. Status and cultivating condition of cells have relationship with sensitivity and accuracy of rotavirus titration.