| 赵慧,李娟,邵铭,刘书珍,徐康维,李长贵.抗H7N9流感病毒中和抗体快速检测方法的建立及应用[J].中国药事,2019,33(1):56-60 |
| 抗H7N9流感病毒中和抗体快速检测方法的建立及应用 |
| Establishment and Application of Rapid Detection Method for Neutralizing Antibody Against Influenza A (H7N9) Virus |
| 投稿时间：2018-01-28 |
| DOI：10.16153/j.1002-7777.2019.01.010 |
| 中文关键词: H7N9流感病毒 中和抗体 微量病毒中和法 血凝抑制试验 |
| 英文关键词: influenza A (H7N9) virus neutralizing antibody micro-virus neutralization method hemagglutination inhibition assay |
| 基金项目:重大新药创制科技重大专项（编号2015ZX09101044）；中国食品药品检定研究院学科带头人培养基金（编号2015X7） |
| 作者 | 单位 | E-mail | | 赵慧 | 中国食品药品检定研究院, 卫生部生物技术产品检定方法及其标准化重点实验室, 北京 102629 | | | 李娟 | 中国食品药品检定研究院, 卫生部生物技术产品检定方法及其标准化重点实验室, 北京 102629 | | | 邵铭 | 中国食品药品检定研究院, 卫生部生物技术产品检定方法及其标准化重点实验室, 北京 102629 | | | 刘书珍 | 中国食品药品检定研究院, 卫生部生物技术产品检定方法及其标准化重点实验室, 北京 102629 | | | 徐康维 | 中国食品药品检定研究院, 卫生部生物技术产品检定方法及其标准化重点实验室, 北京 102629 | | | 李长贵 | 中国食品药品检定研究院, 卫生部生物技术产品检定方法及其标准化重点实验室, 北京 102629 | changguili@aliyun.com |
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| 中文摘要: |
| 目的：对建立的抗H7N9流感病毒中和抗体快速检测方法进行方法学验证及初步应用。方法：分别采用不同代次细胞对高、中、低不同滴度的阳性血清进行多次平行检测，考察细胞代次对检验结果的影响；采用NIBSC提供的参考品对方法学的特异性进行验证；同时应用抗H7N9的阳性血清检测进一步评估方法的准确性和精密性；采用ELISA-MNT法和血凝抑制（hemagglutination inhibition，HI）试验分别接种H7N9灭活流感疫苗免疫的小鼠血清样本20份，分析两种方法检测结果的相关性。结果：采用ELISA-MNT中和法，使用不同代次MDCK细胞（25、30和35代）检测相同的血清样本的中和抗体滴度结果相同；该方法只对羊抗H7N9的血清具有较高保护力，对其他血清基本没有交叉反应；该方法准确性良好；该方法组间、组内精密性的平均变异系数分别为4%和11%。该方法测定H7N9型流感疫苗免疫后的小鼠血清抗体效价，其结果与HI抗体的相关系数为0.61，表明两种方法的检测结果之间呈良好的正相关性。结论：建立的微量病毒中和法能够满足H7N9流感病毒中和抗体效价检测的要求，可用于H7N9新型大流行流感疫苗的免疫评价。 |
| 英文摘要: |
| Objective:To carry out methodological validation and preliminary application of the rapid detection method for neutralizing antibody against influenza A(H7N9) virus. Methods:Different generations of positive cells were used to perform multiple parallel tests on high, medium and low titers of positive serums, and the effects of cell generation on the test results were investigated. Specificity of the method was verified by the reference material provide by NIBSC. Accuracy and precision of the method were evaluated by anti-H7N9 virus positive serums. ELISA-MNT and hemagglutination inhibition (HI) assay were separately used to test the antibody titers from mouse serums immunized with inactivated influenza H7N9 vaccine and the correlation of the two assays was evaluated. Results:The neutralizing antibody titers of the same serum samples were identical by using the ELISA-MNT neutralization method and different generations of MDCK cells (the 25th, 30th and 35th generation). The method only had high protection against the serum of sheep anti-H7N9 and had no cross reaction with other serums. The method had good accuracy and average coefficient variations of intra-assay and inter-assay were 4% and 11.2%, respectively. The correlation coefficient between the antibody titers from mice immunized with H7N9 influenza vaccine measured by the ELISA-MNT assay that obtained from the HI assay was 0.61, indicating that there was a good positive correlation between the detection results of the two methods. Conclusion:The established micro-virus neutralization method can meet the requirement of H7N9 influenza virus neutralizing antibody titer detection, and can be used for immunological evaluation of vaccine for H7N9 new pandemic influenza. |
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