Applicability validation and application of Digoxigenin-labeled DNA probe in detection of residual DNA by hybridization in Vero cell

摘要

Objective Objective To detect the residual DNA quantity in Vero cell contained in rabies vaccine( Vero cell) for human use(simplified as rabies vaccine )by Digoxigenin-labeled DNA probe,and the applicability of the method was verified and applied. Methods The Digoxigenin-labeled DNA probe was carried out in verification of the specificity,sensi-tivity,repeatability and stability for detecting the residual DNA quantity in Vero cell contained in rabies vaccine,and the method was applied to detect residual DNA quantity in Vero cell of 3 batches of rabies vaccine. Results The labeling effi-ciency could reach 0. 1 pg. The verification result showed that there was no hybridazation between the labeled probe and heterologous DNA,and the minimal detective limit reached 1. 0 pg and the detective sensitivity was kept in 1. 0 pg after the labeled probe stored at-20℃ for 7 months. The results conformed to the quality control standards published in《Pharmar-copoeia of the People’ s Republic of China》in detecting residual DNA quantity in Vero cell in 3 batches of rabies vaccine. Conclusion The dot hybridization method is specific,sensitive,stable and well repeatable,it is used in detecting residual DNA quantity in Vero cell contained in rabies vaccine. As a result,the method is suitable in production and quantity control of rabies vaccine,it has a helpful effect in preparation of the other viral vaccines by using Vero cell as a substrate.

关键词