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根据A/Anhui/1/2013(H7N9)病毒株HA和NA基因序列研究H7N9禽流感病毒的结构并制备相应的假病毒，并对此假病毒进行初步验证。将H7N9病毒株的HA和NA基因序列经公司优化合成后，经过接菌、提取质粒、假病毒构建等步骤来制备假病毒，并通过假病毒感染实验、透射电镜以及血清中和实验来验证假病毒制备是否成功。成功制备滴度均超过标准值104的A/Anhui/1/2013(H7N9)假病毒，在电镜下可观察到典型的流感病毒形态，并进一步使用本科室保存的29份血清样本（含4份H7N9阳性血清样本）、国家流感中心制备的四种不同亚型免疫组分（H1型、H3型、BV型和BY型）的抗血清以及商品化H7N9特异性抗体进行中和试验，结果显示4份阳性血清中和活性明显高于25份阴性血清，四种不同亚型抗血清的中和效率均低于40%，且商品化H7N9特异性抗体能够与其有效结合。本研究成功构建了H7N9假病毒并有效筛选出阳性血清，且具有良好的特异性，为流感大流行或疫苗免疫后人群血清学监测建立了科学有效的技术手段。
Abstract：According to the HA and NA gene sequences of A/Anhui/1/2013(H7N9) virus strain, the structure of H7N9 avian influenza virus was studied, and the corresponding pseudovirus was prepared and preliminarily verified. After the HA and NA gene sequences of H7N9 virus strain were optimized and synthesized by the company, pseudovirus was prepared by inoculation, plasmid extraction, pseudovirus construction and other steps, and the success of pseudovirus preparation was verified by pseudovirus infection experiment, transmission electron microscope and serum neutralization assay. A/Anhui/1/2013(H7N9) pseudoviruses with titers exceeding the standard value of 104 were successfully prepared, and typical influenza virus morphology could be observed under electron microscope. Furthermore, 29 serum samples(including 4 H7N9 positive serum samples) preserved by our department, antiserum against four different subtypes of immune components(H1, H3, BV and BY) prepared by National Influenza Center and commercial H7N9 specific antibody were used for neutralization assay. Results showed that the neutralization activity of four positive sera was significantly higher than that of 25 negative sera, and the neutralization efficiency of four different subtypes of antiserum was lower than 40%, and commercial H7N9 specific antibody could effectively bind with them. In this study, the H7N9 pseudovirus was successfully constructed and the positive serum was effectively screened, with good specificity, which established a scientific and effective technical means for serological monitoring of people after influenza pandemic or vaccine immunization.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.004381
中图分类号:R373
引用信息:
[1]胡敏昊,王玮,吴小珉,等.H7N9亚型禽流感假病毒的制备与验证[J].病毒学报,2023,39(05):1227-1234.DOI:10.13242/j.cnki.bingduxuebao.004381.
基金信息:
2021年度安徽省卫生健康科研项目（项目号：AHWJ2021a030）,题目：安徽省候鸟迁徙活动对本地禽流感病毒基因重配及传播影响研究; 科技部国家重点研发计划政府间国际科技创新合作项目（项目号：2022YFE0110100）,题目：聚集性场所空气中多种病毒在线探测与预警关键技术研究~~
2023-03-16
2023
2023-05-13
2023-08-30
2023
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2023-08-31
2023-08-31
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