Persistence of infectious SARS-CoV-2 particles for up to 37 days in patients with mild COVID-19.
Persistence of infectious SARS-CoV-2 particles for up to 37 days in patients with mild COVID-19.
复制标题
在轻度COVID-19患者中，传染性SARS-CoV-2颗粒持续长达37天。
作者：
Zahn T;Mhedhbi I;Hein S;Raupach J;Miskey C;Husria Y;Bayanga K;Bartel D;Vieths S;Ivics Z;Oberle D;Keller-Stanislawski B;Herrlein ML;Maier TJ;Hildt E
患有COVID-19的人通常在诊断后14天被认为是非传染性的，如果症状消失至少48小时。我们描述了三个病人谁独立获得他们的感染。这三名患者经历了轻度COVID-19，并在10天内完全康复，但在咽深部样本中保持PCR阳性至少38天。我们尝试从咽拭子中分离病毒，以调查这些患者是否仍携带传染性病毒。感染性病毒在Vero E6细胞中扩增，并通过电子显微镜和WGS进行表征。用ELISA和肽芯片检测免疫应答。在所有三种情况下，在Vero E6细胞中分离并扩增了感染性和复制能力病毒。通过RT-PCR和免疫荧光显微镜检测病毒复制。电子显微镜证实了完整的SARS-CoV-2颗粒的形成。为了进行更详细的分析，通过全基因组测序（WGS）对所有三种分离株进行了表征。序列数据显示，这些分离株属于20 A或20 C进化枝，并且在可能与建立长期感染相关的其他突变中鉴定出ORF 8中的两个突变。与从轻度COVID-19完全恢复的患者相比，体液免疫应答的表征显示，长期感染患者缺乏与受体结合结构域（RBD）的顺序表位结合的抗体。因此，一小部分COVID-19患者表现出长期传染性，在14天后终止隔离期，而不进行基于PCR的检测，应该重新考虑。这项研究显示，三名长期SARS-CoV-2 PCR阳性患者持续产生感染性病毒长达38天，没有持续症状。与从轻度COVID-19中完全恢复的患者相比，患者显示出低抗体滴度和IgG对刺突蛋白序列表位的结合减少。WGS显示分离株属于20 A或20 C分支，病毒根除可能受到ORF 8中两个突变的影响。缩写：COVID-19，冠状病毒病2019; M，膜蛋白; ORF，开放阅读框; qPCR，定量聚合酶链反应; S，刺突蛋白; SARS-CoV-2，严重急性呼吸综合征冠状病毒2型; WGS，全基因组测序
People suffering from COVID‐19 are typically considered non‐infectious 14 days after diagnosis if symptoms have disappeared for at least 48 h. We describe three patients who independently acquired their infection. These three patients experienced mild COVID‐19 and completely recovered symptomatically within 10 days, but remained PCR‐positive in deep pharyngeal samples for at least 38 days. We attempted to isolate virus from pharyngeal swabs to investigate whether these patients still carried infectious virus. Infectious virus was amplified in Vero E6 cells and characterized by electron microscopy and WGS. The immune response was investigated by ELISA and peptide arrays. In all three cases, infectious and replication‐competent virus was isolated and amplified in Vero E6 cells. Virus replication was detected by RT‐PCR and immunofluorescence microscopy. Electron microscopy confirmed the formation of intact SARS‐CoV‐2 particles. For a more detailed analysis, all three isolates were characterized by whole‐genome sequencing (WGS). The sequence data revealed that the isolates belonged to the 20A or 20C clade, and two mutations in ORF8 were identified among other mutations that could be relevant for establishing a long‐term infection. Characterization of the humoral immune response in comparison to patients that had fully recovered from mild COVID‐19 revealed a lack of antibodies binding to sequential epitopes of the receptor‐binding domain (RBD) for the long‐term infected patients. Thus, a small portion of COVID‐19 patients displays long‐term infectivity and termination of quarantine periods after 14 days, without PCR‐based testing, should be reconsidered critically. This study shows three long‐term SARS‐CoV‐2 PCR‐positive patients persistently producing infectious virus for up to 38 days without lasting symptoms. Patients displayed low antibody titers and decreased binding of IgG against sequential epitopes of spike protein in comparison to patients that fully recovered from mild COVID‐19. WGS revealed that isolates belong to the 20A or 20C clade and viral eradication might be affected by two mutations in ORF8.Abbreviations: COVID‐19, coronavirus disease 2019; M, membrane protein; ORF, open reading frame; qPCR, quantitative polymerase chain reactions; S, spike protein; SARS‐CoV‐2, severe acute respiratory syndrome coronavirus 2; WGS, whole genome sequencing
登录
查看更多内容
影响因子：
64.8
作者：
Plante JA;Liu Y;Liu J;Xia H;Johnson BA;Lokugamage KG;Zhang X;Muruato AE;Zou J;Fontes-Garfias CR;Mirchandani D;Scharton D;Bilello JP;Ku Z;An Z;Kalveram B;Freiberg AN;Menachery VD;Xie X;Plante KS;Weaver SC;Shi PY
通讯作者：
Shi PY
影响因子：
28.3
作者：
Gorbalenya, Alexander E.;Baker, Susan C.;Ziebuhr, John
通讯作者：
Ziebuhr, John
影响因子：
168.9
作者：
Huang, Chaolin;Wang, Yeming;Cao, Bin
通讯作者：
Cao, Bin
影响因子：
82.9
作者：
Amanat F;Stadlbauer D;Strohmeier S;Nguyen THO;Chromikova V;McMahon M;Jiang K;Arunkumar GA;Jurczyszak D;Polanco J;Bermudez-Gonzalez M;Kleiner G;Aydillo T;Miorin L;Fierer DS;Lugo LA;Kojic EM;Stoever J;Liu STH;Cunningham-Rundles C;Felgner PL;Moran T;García-Sastre A;Caplivski D;Cheng AC;Kedzierska K;Vapalahti O;Hepojoki JM;Simon V;Krammer F
通讯作者：
Krammer F
DOI：
10.1007/s10096-020-03913-9
发表时间：
2020-04-27
影响因子：
4.5
作者：
La Scola, Bernard;Le Bideau, Marion;Raoult, Didier
通讯作者：
Raoult, Didier