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[Preliminary study on RT-PCR-ELISA method for virus titer testing of live attenuated hepatitis A vaccine]
Wen Qian1, Yue-qing Chen, Yan Hong
1Institute of Viral Diseases, Zhejiang Academy of Medical Sciences, Hangzhou 310013, China.
Objective:
To evaluate the RT-PCR-ELISA method applied for testing live attenuated hepatitis A vaccine titer.
Methods:
A solid phase hybridization-enzyme colorimetric detection method was used for detecting specific nucleic acid. Primer labeled with biotin was used to amplify viral gene fragment, then the product was quickly hybridized with the specific probe covalently coupled on DNA-binding microplate wells. Finally, peroxidase-labeled streptavidin was used in colorimetric detection. The results were judged by reading A value. Eleven batches of live attenuated hepatitis A vaccine titer were tested by this method. The results were compared with that of routine cell culture method (CCID50).
Results:
The sensitivity was similar to routine cell culture method (P>0.05). This method was convenient, fast and specific.
Conclusion:
CCID50 method may be replaced by the RT-PCR-ELISA method in evaluating the titer of live attenuated hepatitis A vaccine.
