A line immunoassay utilizing recombinant nucleocapsid proteins for detection of antibodies to human coronaviruses

Diagnostic Microbiology and Infectious Disease
Christian LehmannPetra Lindner

Abstract

Most coronaviruses infecting humans cause mild diseases, whereas severe acute respiratory syndrome (SARS)-associated coronavirus is an extremely dangerous pathogen. Here, we report the development of a serologic assay for detection of antibodies to human coronaviruses (HCoVs) based on recombinant nucleocapsid (N) proteins of all known pathogenic strains (229E, NL63, OC43, HKU1, SARS). The novel immunoassay is highly useful for epidemiologic surveys, where use of nucleic acid diagnostics often is limited. Purified recombinant antigens were immobilized on nitrocellulose membranes and applied in a line immunoassay, which allows rapid detection of antibodies to 5 different HCoVs in a single experiment. For assay evaluation, serum samples from persons infected with 229E or OC43 (acute/convalescent), recovered SARS patients and healthy donors were analyzed. Screening for nucleocapsid (N)-specific immunoglobulin G (IgG) in convalescent sera reached 100% sensitivity. With this new technique, we found that recently identified NL63 and HKU1 contribute significantly to the overall spectrum of coronavirus infections. Possibly, cross-reactive antibody responses were observed using 229E and OC43 serum pairs. However, the potential of this as...Continue Reading

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Citations

Jun 9, 2009·The Pediatric Infectious Disease Journal·Christian LehmannStephan Gerling
May 23, 2008·Journal of Clinical Microbiology·Ronald DijkmanLia van der Hoek
Dec 23, 2011·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Ronald DijkmanLia van der Hoek
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Apr 17, 2020·The Journal of Infectious Diseases·Sema NickbakhshPablo R Murcia
Apr 9, 2020·Frontiers in Microbiology·Ning WangLanying Du
Sep 16, 2020·Nature Medicine·Arthur W D EdridgeLia van der Hoek

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Datasets Mentioned

BETA
DQ243939
AY585228
DQ778921
DQ437631

Methods Mentioned

BETA
gel
electrophoresis
gel filtration
enzyme-linked immunosorbent assay
PCR

Software Mentioned

GCG Wisconsin Package

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