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 RT PCR 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测。
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 detectionThe results were judged by reading A valueEleven batches of live attenuated hepatitis A vaccine titer were tested by this method The results were compared with that of routine cell culture method (CCID 50 ) Results The sensitivity was similar to routine cell culture method (P>005) This method was convenient, fast and specific Conclusion CCID 50 method may be replaced by the RT-PCR-ELISA method in evaluating the titer of live attenuated hepatitis A vaccine
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
北大核心
2004年第3期261-264,共4页
Chinese Journal of Experimental and Clinical Virology
