| RT-PCR-ELISA方法检测甲肝减毒活疫苗病毒滴度的初步研究 |
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| 引用本文： | 钱汶,陈悦青,洪艳,唐彩华,周康凤,庄昉成,陈勇. RT-PCR-ELISA方法检测甲肝减毒活疫苗病毒滴度的初步研究[J]. 中华实验和临床病毒学杂志, 2004, 18(3): 261-264 |
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| 作者姓名： | 钱汶 陈悦青 洪艳 唐彩华 周康凤 庄昉成 陈勇 |
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| 作者单位： | 1. 310013,杭州,浙江省医学科学院病毒病研究所检定室
2. 310013,杭州,浙江省医学科学院生物工程所 |
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| 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测。
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| 关 键 词： | 甲肝减毒活疫苗 病毒滴度 RT-PCR 细胞培养法 ELISA方法 初步研究 亲和素 杂交 增产 引物 |
| 修稿时间： | 2004-06-10 |
Preliminary study on RT-PCR-ELISA method for virus titer testing of live attenuated hepatitis A vaccine |
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| QIAN Wen ,CHEN Yue-qing,HONG Yan,TANG Cai-hua,ZHOU Kang-feng,ZHUANG Fang-cheng,CHEN Yong Institute of Viral Diseases,Zhejiang Academy of Medical Sciences,Hangzhou ,China. Preliminary study on RT-PCR-ELISA method for virus titer testing of live attenuated hepatitis A vaccine[J]. Chinese journal of experimental and clinical virology, 2004, 18(3): 261-264 |
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| Authors: | QIAN Wen CHEN Yue-qing HONG Yan TANG Cai-hua ZHOU Kang-feng ZHUANG Fang-cheng CHEN Yong Institute of Viral Diseases Zhejiang Academy of Medical Sciences Hangzhou China |
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| Affiliation: | QIAN Wen *,CHEN Yue-qing,HONG Yan,TANG Cai-hua,ZHOU Kang-feng,ZHUANG Fang-cheng,CHEN Yong * Institute of Viral Diseases,Zhejiang Academy of Medical Sciences,Hangzhou 310013,China |
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| 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 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 |
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