Science Translational Medicine ( IF 17.1 ) Pub Date : 2020-09-02 , DOI: 10.1126/scitranslmed.abc3103 Ludivine Grzelak 1, 2, 3 , Sarah Temmam 4 , Cyril Planchais 5 , Caroline Demeret 6, 7 , Laura Tondeur 8 , Christèle Huon 4 , Florence Guivel-Benhassine 1, 2 , Isabelle Staropoli 1, 2 , Maxime Chazal 9 , Jeremy Dufloo 1, 2, 3 , Delphine Planas 1, 2 , Julian Buchrieser 1, 2 , Maaran Michael Rajah 1, 2, 3 , Remy Robinot 1, 2 , Françoise Porrot 1, 2 , Mélanie Albert 6, 7, 10 , Kuang-Yu Chen 11 , Bernadette Crescenzo-Chaigne 6, 7 , Flora Donati 6, 7, 10 , François Anna 12 , Philippe Souque 13 , Marion Gransagne 14 , Jacques Bellalou 15 , Mireille Nowakowski 15 , Marija Backovic 16 , Lila Bouadma 17, 18 , Lucie Le Fevre 18 , Quentin Le Hingrat 17, 19 , Diane Descamps 17, 19 , Annabelle Pourbaix 20 , Cédric Laouénan 17, 21 , Jade Ghosn 17, 20 , Yazdan Yazdanpanah 17, 20 , Camille Besombes 8 , Nathalie Jolly 22 , Sandrine Pellerin-Fernandes 22 , Olivia Cheny 22 , Marie-Noëlle Ungeheuer 22 , Guillaume Mellon 23 , Pascal Morel 24 , Simon Rolland 25, 26 , Felix A Rey 16 , Sylvie Behillil 6, 7, 10 , Vincent Enouf 6, 7, 10 , Audrey Lemaitre 27 , Marie-Aude Créach 28, 29 , Stephane Petres 15 , Nicolas Escriou 14 , Pierre Charneau 12, 13 , Arnaud Fontanet 8, 30 , Bruno Hoen 31 , Timothée Bruel 1, 2 , Marc Eloit 4, 32 , Hugo Mouquet 5 , Olivier Schwartz 1, 2 , Sylvie van der Werf 6, 7, 10
It is of paramount importance to evaluate the prevalence of both asymptomatic and symptomatic cases of SARS-CoV-2 infection and their differing antibody response profiles. Here, we performed a pilot study of four serological assays to assess the amounts of anti–SARS-CoV-2 antibodies in serum samples obtained from 491 healthy individuals before the SARS-CoV-2 pandemic, 51 individuals hospitalized with COVID-19, 209 suspected cases of COVID-19 with mild symptoms, and 200 healthy blood donors. We used two ELISA assays that recognized the full-length nucleoprotein (N) or trimeric spike (S) protein ectodomain of SARS-CoV-2. In addition, we developed the S-Flow assay that recognized the S protein expressed at the cell surface using flow cytometry, and the luciferase immunoprecipitation system (LIPS) assay that recognized diverse SARS-CoV-2 antigens including the S1 domain and the carboxyl-terminal domain of N by immunoprecipitation. We obtained similar results with the four serological assays. Differences in sensitivity were attributed to the technique and the antigen used. High anti–SARS-CoV-2 antibody titers were associated with neutralization activity, which was assessed using infectious SARS-CoV-2 or lentiviral-S pseudotype virus. In hospitalized patients with COVID-19, seroconversion and virus neutralization occurred between 5 and 14 days after symptom onset, confirming previous studies. Seropositivity was detected in 32% of mildly symptomatic individuals within 15 days of symptom onset and in 3% of healthy blood donors. The four antibody assays that we used enabled a broad evaluation of SARS-CoV-2 seroprevalence and antibody profiling in different subpopulations within one region.
中文翻译：
四种血清学检测方法的比较，用于检测来自不同人群的人血清样品中的抗SARS-CoV-2抗体。
评估SARS-CoV-2感染的无症状和有症状病例的患病率及其不同的抗体反应谱至关重要。在这里，我们对四种血清学检测方法进行了初步研究，以评估从SARS-CoV-2大流行之前的491名健康个体（其中51例因COVID-19住院的个体）中获得的血清样本中抗SARS-CoV-2抗体的量，209疑似COVID-19病例，症状轻微，有200名健康献血者。我们使用了两种ELISA分析法来识别SARS-CoV-2的全长核蛋白（N）或三聚体刺突（S）蛋白胞外域。此外，我们开发了S-Flow检测方法，该方法可使用流式细胞仪识别在细胞表面表达的S蛋白，荧光素酶免疫沉淀系统（LIPS）测定法通过免疫沉淀法识别了多种SARS-CoV-2抗原，包括N的S1结构域和羧基末端结构域。我们通过四种血清学测定获得了相似的结果。敏感性的差异归因于技术和所使用的抗原。抗SARS-CoV-2高抗体滴度与中和活性有关，中和活性使用感染性SARS-CoV-2或慢病毒S假型病毒进行评估。在住院的COVID-19患者中，症状发作后5到14天之间发生了血清转化和病毒中和，这证实了以前的研究。在症状发作后15天内，有32％的轻度有症状个体和3％的健康献血者中发现了血清阳性。
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