A pseudovirus-based platform to measure neutralizing antibodies in Mexico using SARS-CoV-2 as proof-of-concept.
A pseudovirus-based platform to measure neutralizing antibodies in Mexico using SARS-CoV-2 as proof-of-concept.
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DOI：
10.1038/s41598-022-22921-7
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发表时间：
2022-10-26
影响因子：
4.6
通讯作者：
中科院分区：
文献类型：
--
作者：
评估功能性抗病毒免疫反应的金标准方法是滴度针对病毒病原体的中和抗体（nab）。这在过去是使用病毒介导感染的体外试验来进行的，这需要BSL-3设施。由于在包括墨西哥在内的拉丁美洲国家缺乏这些工具，因此使用功能测定法在当地获得的关于病毒性病原体NAb的信息很少。对活病毒使用BSL-3试验的另一种解决方案是对非复制性假病毒使用bsl -2安全试验。假病毒颗粒可以被设计成在其表面显示选定病原体的进入蛋白，并在转导时将报告基因传递到靶细胞。在这里，我们全面描述了在墨西哥首次开发的基于伪型慢病毒颗粒生产的BSL-2安全nabs测量功能分析。作为概念验证，该检测方法基于纳米荧光素酶介导的靶细胞的发光测量，这些细胞被SARS-CoV-2刺状假慢病毒颗粒转导。我们将优化后的方法应用于BSL-2设施中，检测了65份血清样本的nab，结果表明，该方法的灵敏度为100%，特异性为86.6%，准确度为96%。总的来说，这是墨西哥开发的基于BSL-2安全假病毒的功能测定法的第一份报告，也是在资源有限的情况下用功能测定法测量nab所需的基础方法。
The gold-standard method to evaluate a functional antiviral immune response is to titer neutralizing antibodies (NAbs) against a viral pathogen. This is historically performed using an in vitro assay of virus-mediated infection, which requires BSL-3 facilities. As these are insufficient in Latin American countries, including Mexico, scant information is obtained locally about viral pathogens NAb, using a functional assay. An alternative solution to using a BSL-3 assay with live virus is to use a BSL-2-safe assay with a non-replicative pseudovirus. Pseudoviral particles can be engineered to display a selected pathogen’s entry protein on their surface, and to deliver a reporter gene into target cells upon transduction. Here we comprehensively describe the first development of a BSL-2 safe NAbs-measuring functional assay in Mexico, based on the production of pseudotyped lentiviral particles. As proof-of-concept, the assay is based on Nanoluc luciferase-mediated luminescence measurements from target cells transduced with SARS-CoV-2 Spike-pseudotyped lentiviral particles. We applied the optimized assay in a BSL-2 facility to measure NAbs in 65 serum samples, which evidenced the assay with 100% sensitivity, 86.6% specificity and 96% accuracy. Overall, this is the first report of a BSL-2 safe pseudovirus-based functional assay developed in Mexico to measure NAbs, and a cornerstone methodology necessary to measure NAbs with a functional assay in limited resources settings.
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影响因子：
5.4
作者：
Heurich, Adeline;Hofmann-Winkler, Heike;Poehlmann, Stefan
通讯作者：
Poehlmann, Stefan
影响因子：
9.2
作者：
Chen M;Zhang XE
通讯作者：
Zhang XE
影响因子：
2.7
作者：
Hattermann, K;Müller, MA;Niedrig, M
通讯作者：
Niedrig, M
DOI：
10.1074/jbc.m505111200
发表时间：
2005-08-26
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Lambert DW;Yarski M;Warner FJ;Thornhill P;Parkin ET;Smith AI;Hooper NM;Turner AJ
通讯作者：
Turner AJ
影响因子：
9.1
作者：
Hicks J;Klumpp-Thomas C;Kalish H;Shunmugavel A;Mehalko J;Denson JP;Snead KR;Drew M;Corbett KS;Graham BS;Hall MD;Memoli MJ;Esposito D;Sadtler K
通讯作者：
Sadtler K