RT Journal Article
SR Electronic
T1 Highly sensitive and specific multiplex antibody assays to quantify immunoglobulins M, A and G against SARS-CoV-2 antigens
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.06.11.147363
DO 10.1101/2020.06.11.147363
A1 Carlota Dobaño
A1 Marta Vidal
A1 Rebeca Santano
A1 Alfons Jiménez
A1 Jordi Chi
A1 Diana Barrios
A1 Gemma Ruiz-Olalla
A1 Natalia Rodrigo Melero
A1 Carlo Carolis
A1 Daniel Parras
A1 Pau Serra
A1 Paula Martínez de Aguirre
A1 Francisco Carmona-Torre
A1 Gabriel Reina
A1 Pere Santamaria
A1 Alfredo Mayor
A1 Alberto García-Basteiro
A1 Luis Izquierdo
A1 Ruth Aguilar
A1 Gemma Moncunill
YR 2020
UL http://biorxiv.org/content/early/2020/06/12/2020.06.11.147363.abstract
AB Reliable serological tests are required to determine the prevalence of antibodies against SARS-CoV-2 antigens and to characterise immunity to the disease in order to address key knowledge gaps in the context of the COVID-19 pandemic. Quantitative suspension array technology (qSAT) assays based on the xMAP Luminex platform overcome the limitations of rapid diagnostic tests and ELISA with their higher precision, dynamic range, throughput, miniaturization, cost-efficacy and multiplexing capacity. We developed three qSAT assays to detect IgM, IgA and IgG to a panel of eight SARS-CoV-2 antigens including spike (S), nucleoprotein (N) and membrane (M) protein constructs. The assays were optimized to minimize processing time and maximize signal to noise ratio. We evaluated the performance of the assays using 128 plasmas obtained before the COVID-19 pandemic (negative controls) and 115 plasmas from individuals with SARS-CoV-2 diagnosis (positive controls), of whom 8 were asymptomatic, 58 had mild symptoms and 49 were hospitalized. Pre-existing IgG antibodies recognizing N, M and S2 proteins were detected in negative controls suggestive of cross-reactive to common cold coronaviruses. The best performing antibody isotype/antigen signatures had specificities of 100% and sensitivities of 94.94% at ≥14 days since the onset of symptoms and 96.08% at ≥21 days since the onset of symptoms, with AUC of 0.992 and 0.999, respectively. Combining multiple antibody markers as assessed by qSAT assays has the highest efficiency, breadth and versatility to accurately detect low-level antibody responses for obtaining reliable data on prevalence of exposure to novel pathogens in a population. Our assays will allow gaining insights into antibody correlates of immunity required for vaccine development to combat pandemics like the COVID-19.Competing Interest StatementThe authors have declared no competing interest.