A line immunoassay utilizing recombinant nucleocapsid proteins for detection of antibodies to human coronaviruses
Identifieur interne : 000302 ( PascalFrancis/Checkpoint ); précédent : 000301; suivant : 000303A 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.
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- Pascal (Inist)
- Wicri :
- topic : Homme.
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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.
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<front><div type="abstract" xml:lang="en">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.</div>
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</fC07>
<fC07 i1="01" i2="X" l="ENG"><s0>Coronaviridae</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="01" i2="X" l="SPA"><s0>Coronaviridae</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="02" i2="X" l="FRE"><s0>Nidovirales</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="02" i2="X" l="ENG"><s0>Nidovirales</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="02" i2="X" l="SPA"><s0>Nidovirales</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="03" i2="X" l="FRE"><s0>Virus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="03" i2="X" l="ENG"><s0>Virus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="03" i2="X" l="SPA"><s0>Virus</s0>
<s2>NW</s2>
</fC07>
<fC07 i1="04" i2="X" l="FRE"><s0>Epidémiologie</s0>
<s5>13</s5>
</fC07>
<fC07 i1="04" i2="X" l="ENG"><s0>Epidemiology</s0>
<s5>13</s5>
</fC07>
<fC07 i1="04" i2="X" l="SPA"><s0>Epidemiología</s0>
<s5>13</s5>
</fC07>
<fN21><s1>154</s1>
</fN21>
<fN44 i1="01"><s1>OTO</s1>
</fN44>
<fN82><s1>OTO</s1>
</fN82>
</pA>
</standard>
</inist>
<affiliations><list><country><li>Allemagne</li>
<li>République populaire de Chine</li>
</country>
<settlement><li>Pékin</li>
</settlement>
</list>
<tree><country name="Allemagne"><noRegion><name sortKey="Lehmann, Christian" sort="Lehmann, Christian" uniqKey="Lehmann C" first="Christian" last="Lehmann">Christian Lehmann</name>
</noRegion>
<name sortKey="Lindner, Petra" sort="Lindner, Petra" uniqKey="Lindner P" first="Petra" last="Lindner">Petra Lindner</name>
<name sortKey="Motz, Manfred" sort="Motz, Manfred" uniqKey="Motz M" first="Manfred" last="Motz">Manfred Motz</name>
<name sortKey="Wolf, Hans" sort="Wolf, Hans" uniqKey="Wolf H" first="Hans" last="Wolf">Hans Wolf</name>
</country>
<country name="République populaire de Chine"><noRegion><name sortKey="Jianguo Xu" sort="Jianguo Xu" uniqKey="Jianguo Xu" last="Jianguo Xu">JIANGUO XU</name>
</noRegion>
<name sortKey="Quanbi Zhao" sort="Quanbi Zhao" uniqKey="Quanbi Zhao" last="Quanbi Zhao">QUANBI ZHAO</name>
<name sortKey="Yiming Shao" sort="Yiming Shao" uniqKey="Yiming Shao" last="Yiming Shao">YIMING SHAO</name>
</country>
</tree>
</affiliations>
</record>
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