重组戊型肝炎疫苗宿主细胞残余DNA荧光染色法检测方法的建立和验证

Method establishment and verification for the detection of host cell DNA residue in recombinant hepatitis E vaccine by fluorescence staining

目的建立检测重组戊型肝炎疫苗宿主DNA残留量的荧光染色法,并进行方法验证。方法采用λDNA作为标准品建立标准曲线,重组戊型肝炎疫苗纯化后原液作为样品,进行线性、准确度以及精密度的验证。结果该方法在1.250~80.000 ng/ml范围内有良好的线性,决定系数大于0.99;准确性验证中DNA残留量加标回收率在90.00%~110.00%之间,且加标回收率相对标准偏差均小于15.00%。重复性验证和中间精密度验证的相对标准偏差分别为6.62%和10.30%,均小于15.00%。结论该方法快速、灵敏、准确,可以检测重组戊型肝炎疫苗宿主DNA残留量。

Objective To establish and verify a fluorescence staining method for the residual DNA content in recombinant hepatitis E vaccine(HepEV).MethodsλDNA was used as the standard to establish the standard curve,and purified recombinant HepEV bulk was used as the sample to verify the linearity,accuracy and precision.Results The method had good linearity in the range of 1.250-80.000 ng/ml,and the coefficient of determination was greater than 0.99.In the accuracy verification,the recovery rate of DNA residue was between 90%and 110%,and the relative standard deviation(RSD)of the recovery rate was less than 15%.At the same time,the RSDs of repeatability verification and intermediate precision verification were 6.62%and 10.30%,respectively,both less than 15%.Conclusion This method is rapid,sensitive and accurate,and can detect the residual host DNA content in recombinant HepEV.