Cross-reactivity of two SARS-CoV-2 serological assays in a malaria-endemic setting.

Steinhardt, Laura C; Ige, Fehintola; Iriemenam, Nnaemeka C; Greby, Stacie M; Hamada, Yohhei; Uwandu, Mabel; Aniedobe, Maureen; Stafford, Kristen A; Abimiku, Alash'le; Mba, Nwando; Agala, Ndidi; Okunoye, Olumide; Mpamugo, Augustine; Swaminathan, Mahesh; Onokevbagbe, Edewede; Olaleye, Temitope; Odoh, Ifeanyichukwu; Marston, Barbara J; Okoye, McPaul; Abubakar, Ibrahim; Rangaka, Molebogeng X; Rogier, Eric; Audu, Rosemary

Background: Accurate SARS-CoV-2 serological assays are critical for COVID-19 serosurveillance. However, previous studies have indicated possible cross-reactivity of these assays, including in malaria-endemic areas.Methods: We tested 213 well-characterized pre-pandemic samples from Nigeria using two SARS-CoV-2 serological assays: Abbott Architect IgG and Euroimmun NCP IgG assay, both targeting SARS-CoV-2 nucleocapsid protein. To assess antibody binding strength, an avidity assay was performed on these samples and on plasma from SARS-CoV-2 PCR-positive persons.Results: Thirteen (6.1%) of 212 samples run on the Abbott assay and 38 (17.8%) of 213 run on the Euroimmun assay were positive. Anti-Plasmodium IgG levels were significantly higher among false-positives for both Abbott and Euroimmun; no association was found with active P. falciparum infection. An avidity assay using various concentratIons of urea wash in the Euroimmun assay reduced loosely-bound IgG: of 37 positive/borderline pre-pandemic samples, 46%, 86%, 89%, and 97% became negative using 2M, 4M, 5M, and 8M urea washes, respectively. The wash slightly reduced avidity of antibodies from SARS-CoV-2 patients within 28 days of PCR confirmation; thereafter avidity increased for all urea concentrations except 8M.Conclusions: This validation found moderate to substantial cross-reactivity on two SARS-CoV-2 serological assays using samples from a malaria-endemic setting. A simple urea wash appeared to alleviate issues of cross-reactivity.