目的: 用试剂盒法建立基于HEK293细胞的腺病毒滴度检测方法,并进行方法学验证,以确定方法的可行性。方法: 用TaKaRa试剂盒检测腺病毒滴度,并对方法的专属性、准确性、精密度、耐用性、线性进行验证。结果: 样品中的佐剂成分、其他型别病毒和异源抗病毒血清均不会对病毒滴度测定结果产生干扰;该方法检测的2个样品病毒滴度,重复性和重现性的RSD均<20%,平均回收率在80%~120%;同一样品不同时间,不同地点检出的结果差异无统计学意义(P均>0.05);试剂盒法检测腺病毒原液和腺病毒成品的病毒滴度的线性范围分别为7.15~9.88 IFU·mL-1和6.43~9.20 IFU·mL-1,在线性范围内,线性回归系数(r)分别为1.000和 0.999 8。结论: 本实验建立的试剂盒法检测腺病毒滴度专属性、准确度、精密度、耐用性、线性良好,是简便、快速、检测误差小的方法,可用于腺病毒滴度的检测。
Objective: To establish an adenovirus titer assay based on HEK293 cells using a kit method and to perform methodological validation to determine the feasibility of the method. Methods: The titer of adenovirus was detected with TaKaRa kit, and the specificity, accuracy, precision, durability and linearity of the method were verified. Results: The adjuvant components, other types of virus and heterologous anti-virus serum in the sample did not interfere with the result of virus titer determination; RSD of repeatability and reproducibility of the two samples detected by this method were both less than 20%, with the average recovery rate between 80% and 120%. There was no statistically significant difference in the results detected at different places at different times and places (P>0. 05). The linear range of viral titers for the detection of adenoviral stock solution and adenovirus finished products by the kit method was 7.15-9.88 IFU·mL-1 and 6.43-9.20 IFU·mL-1, respectively. Within the linear range, the linear regression coefficients (r) were 1.000 and 0.999 8, respectively. Conclusion: The kit method established in this experiment to detect adenovirus titer has good exclusivity, accuracy, precision, durability and linearity. It is a simple, fast method with small detection error, which can be used for the detection of adenovirus titer.
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