Preliminary study on RT-PCR-ELISA method for virus titer testing of live attenuated hepatitis A vaccine.
- Author:
Wen QIAN
1
;
Yue-qing CHEN
;
Yan HONG
;
Cai-hua TANG
;
Kang-feng ZHOU
;
Fang-cheng ZHUANG
;
Yong CHEN
Author Information
- Publication Type:Journal Article
- MeSH: Base Sequence; DNA, Viral; analysis; Enzyme-Linked Immunosorbent Assay; Genes, Viral; Hepatitis A Vaccines; Hepatitis A virus; genetics; Quality Control; Reverse Transcriptase Polymerase Chain Reaction; Sensitivity and Specificity; Vaccines, Attenuated
- From: Chinese Journal of Experimental and Clinical Virology 2004;18(3):261-264
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo evaluate the RT-PCR-ELISA method applied for testing live attenuated hepatitis A vaccine titer.
METHODSA 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).
RESULTSThe sensitivity was similar to routine cell culture method (P>0.05). This method was convenient, fast and specific.
CONCLUSIONCCID50 method may be replaced by the RT-PCR-ELISA method in evaluating the titer of live attenuated hepatitis A vaccine.
