【作者】 钱汶； 陈悦青； 洪艳； 唐彩华； 周康凤； 庄昉成； 陈勇；

【Author】 Qian Wen, Chen Yue-qing, Hong Yan,et al ( Institute of Viral Diseases, Zhejiang Academy of Medical Science, Hangzhou,310013)

【机构】 浙江省医学科学院病毒病研究所； 浙江省医学科学院 生物工程研究所；

【摘要】 目的应用核酸扩增产物测定的固相杂交酶联显色法(RT-PCR-ELISA)检测甲肝减毒活疫苗病毒滴度。方法应用RT-PCR—ELISA,将标记有生物素的寡核苷酸引物所扩增的疫苗病毒基因产物,与微孔反应板上的特异性探针进行快速杂交,通过辣根过氧化物酶标记的链亲和素进行酶联显色,读取吸光度(A值),判断结果。应用此法检测了11批甲肝活疫苗滴度,并与常规细胞培养法(CCID50)比较。结果本方法与细胞培养法的敏感性相仿,具有简便、快速、特异的优点。结论 RTPCR—ELISA法有望代替常规细胞培养法应用于甲肝减毒活疫苗病毒滴度的检浏。更多还原

【Abstract】 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 cova-lently 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.更多还原