A line immunoassay utilizing recombinant nucleocapsid proteins for detection of antibodies to human coronaviruses
Identifieur interne : 003275 ( Main/Merge ); précédent : 003274; suivant : 003276A line immunoassay utilizing recombinant nucleocapsid proteins for detection of antibodies to human coronaviruses
Auteurs : Christian Lehmann [Allemagne] ; Hans Wolf [Allemagne] ; Jianguo Xu [République populaire de Chine] ; Quanbi Zhao [République populaire de Chine] ; Yiming Shao [République populaire de Chine] ; Manfred Motz [Allemagne] ; Petra Lindner [Allemagne]Source :
- Diagnostic Microbiology and Infectious Disease [ 0732-8893 ] ; 2008.
Descripteurs français
- KwdFr :
- Anticorps antiviraux (sang), Coronavirus (immunologie), Dosage immunologique (), Humains, Immunoglobuline G (sang), Infections à coronavirus (diagnostic), Infections à coronavirus (immunologie), Protéines nucléocapside (immunologie), Protéines recombinantes (immunologie), Réactions croisées, Sensibilité et spécificité.
- MESH :
- diagnostic : Infections à coronavirus.
- immunologie : Coronavirus, Infections à coronavirus, Protéines nucléocapside, Protéines recombinantes.
- sang : Anticorps antiviraux, Immunoglobuline G.
- Dosage immunologique, Humains, Réactions croisées, Sensibilité et spécificité.
English descriptors
- KwdEn :
- MESH :
- chemical , blood : Antibodies, Viral, Immunoglobulin G.
- diagnosis : Coronavirus Infections.
- immunology : Coronavirus, Coronavirus Infections, Nucleocapsid Proteins, Recombinant Proteins.
- methods : Immunoassay.
- Cross Reactions, Humans, Sensitivity and Specificity.
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 assay could clearly be demonstrated employing SARS-positive serum samples, where nonspecific binding to nucleocapsids of other HCoVs was not observed. This coronavirus strain-specific line immunoassay represents a powerful tool for serologic diagnostics.
Url:
DOI: 10.1016/j.diagmicrobio.2007.12.002
PubMed: 18191362
PubMed Central: 7127592
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PMC:7127592Le document en format XML
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type="ISSN">0732-8893</idno><idno type="eISSN">1879-0070</idno><imprint><date when="2008">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibodies, Viral (blood)</term><term>Coronavirus (immunology)</term><term>Coronavirus Infections (diagnosis)</term><term>Coronavirus Infections (immunology)</term><term>Cross Reactions</term><term>Humans</term><term>Immunoassay (methods)</term><term>Immunoglobulin G (blood)</term><term>Nucleocapsid Proteins (immunology)</term><term>Recombinant Proteins (immunology)</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Anticorps antiviraux (sang)</term><term>Coronavirus (immunologie)</term><term>Dosage immunologique ()</term><term>Humains</term><term>Immunoglobuline G (sang)</term><term>Infections à coronavirus (diagnostic)</term><term>Infections à coronavirus (immunologie)</term><term>Protéines nucléocapside (immunologie)</term><term>Protéines recombinantes (immunologie)</term><term>Réactions croisées</term><term>Sensibilité et spécificité</term></keywords><keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>Antibodies, Viral</term><term>Immunoglobulin G</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Coronavirus Infections</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Infections à coronavirus</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Coronavirus</term><term>Infections à coronavirus</term><term>Protéines nucléocapside</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Coronavirus</term><term>Coronavirus Infections</term><term>Nucleocapsid Proteins</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Immunoassay</term></keywords><keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>Anticorps antiviraux</term><term>Immunoglobuline G</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Cross Reactions</term><term>Humans</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Dosage immunologique</term><term>Humains</term><term>Réactions croisées</term><term>Sensibilité et spécificité</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>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 assay could clearly be demonstrated employing SARS-positive serum samples, where nonspecific binding to nucleocapsids of other HCoVs was not observed. This coronavirus strain-specific line immunoassay represents a powerful tool for serologic diagnostics.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Arden, K E" uniqKey="Arden K">K.E. Arden</name></author><author><name sortKey="Nissen, M D" uniqKey="Nissen M">M.D. Nissen</name></author><author><name sortKey="Sloots, T P" uniqKey="Sloots T">T.P. Sloots</name></author><author><name sortKey="Sloots, T P" uniqKey="Sloots T">T.P. Sloots</name></author><author><name sortKey="Mackay, I M" uniqKey="Mackay I">I.M. Mackay</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bastien, N" uniqKey="Bastien N">N. 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