[Preliminary study on RT-PCR-ELISA method for virus titer testing of live attenuated hepatitis A vaccine].
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.


CONCLUSIONS
CCID50 method may be replaced by the RT-PCR-ELISA method in evaluating the titer of live attenuated hepatitis A vaccine.
Qian
Wen
W
Institute of Viral Diseases, Zhejiang Academy of Medical Sciences, Hangzhou 310013, China.
Chen
Yue-qing
YQ
Hong
Yan
Y
Tang
Cai-hua
CH
Zhou
Kang-feng
KF
Zhuang
Fang-cheng
FC
Chen
Yong
Y
D001483:Base Sequence; D004279:DNA, Viral / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D005814:Genes, Viral; D022362:Hepatitis A Vaccines; D030041:Hepatitis A virus / Q000235:genetics*; D011786:Quality Control; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014613:Vaccines, Attenuated
D004279:DNA, Viral; D022362:Hepatitis A Vaccines; D014613:Vaccines, Attenuated