Extended Data Fig. 1: Detection of IgG and IgM antibodies in SARS-CoV-2-infected Vero cells. | Nature Medicine

Extended Data Fig. 1: Detection of IgG and IgM antibodies in SARS-CoV-2-infected Vero cells.

From: Breadth of concomitant immune responses prior to patient recovery: a case report of non-severe COVID-19

Extended Data Fig. 1

Immunofluorescence antibody staining for the detection of IgG and IgM bound to SARS-CoV-2-infected vero cells using plasma (diluted 1:20) collected at d7-9 and d20 following onset of symptoms. Immunofluorescence antibody tests for the detection of IgG and IgM were performed using SARS-CoV-2-infected vero cells washed with PBS and methanol/acetone fixed onto glass slides. SARS-CoV-2 was isolated from another COVID patient, and infectivity at cell harvest was ~80%. 10μL of a 1/20 dilution of patient plasma in PBS from days 7, 8, 9 and 20 were incubated on separate wells for 30 mins at 37°C, then washed in PBS and further incubated with 10μL of FITC-conjugated goat anti-human IgG and IgM (Euroimmun, Lűbeck, Germany) before viewing on a EUROStar III Plus fluorescent microscope (Euroimmun). Prior to detection of IgM antibodies, samples were pre-treated with RF-SorboTech (Alere, Rűsselsheim, Germany) to remove IgG antibodies and rheumatoid factors, which may cause false-negative and false-positive IgM results, respectively.

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