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

作者：孙小慧; 刘晓凡; 赵雅静; 马超; 刘鹏; 冯德杰; 李薇; 魏然; 王革

来源：微生物学免疫学进展, 2016, 44(2).

DOI：10.13309/j.cnki.pmi.2016.02.007

• Digoxigenin
• DNA probe
• Dot hybridization
• Vero cell
• Residual DNA quantity

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.