新型冠状病毒假病毒的制备及血浆中和抗体检测Packaging of the SARS-CoV-2 Pseudovirus and Measurement of Neutralizing Antibodies in Serum
伍政宇;罗楚铭;贾婷婷;舒跃龙;
摘要(Abstract):
新型冠状病毒(SARS-CoV-2)引发新型冠状病毒肺炎(COVID-19)全球大流行。由于SARS-CoV-2生物安全管理的要求,高效制备获得高滴度的新型冠状病毒假病毒对基于S蛋白研发疫苗、中和抗体、病毒入侵抑制剂药物以及人群血清学调查等十分重要。本研究基于慢病毒系统,对制备新型冠状病毒假病毒过程中的重要参数进行优化,用Western Blot检测假病毒S蛋白和p24蛋白的表达及假病毒包装情况,用荧光素酶报告系统检测假病毒感染入侵效率。利用制备好的假病毒对恢复期血浆的中和抗体水平进行检测。结果显示骨架质粒与野生型Spike质粒为2∶1比例,在转染后48h收集上清为SARS-CoV-2野生型假病毒包装的最佳条件。与野生型相比,恢复期血浆对四种突变株的中和抗体滴度均降低,对B.1.351株中和能力最弱,B.1.617.2株其次,重型患者恢复期血浆对野生型和突变株的中和抗体滴度高于轻型与普通型患者。本研究优化了新型冠状病毒假病毒包装的实验室条件,评估了COVID-19患者恢复期血浆对野生型及四种突变株的中和抗体水平,提示未来对突变株的免疫逃逸进一步加强监测的重要性。
关键词(KeyWords): 新型冠状病毒;假病毒;中和抗体
基金项目(Foundation): 深圳市科技计划技术攻关重点项目(项目号:JSGG20200225152008136),题目:基于免疫原优化的广谱冠状病毒疫苗研发~~
作者(Authors): 伍政宇;罗楚铭;贾婷婷;舒跃龙;
DOI: 10.13242/j.cnki.bingduxuebao.004126
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