False-positive results in a recombinant severe acute respiratory syndrome-associated coronavirus (SARS-CoV) nucleocapsid-based western blot assay were rectified by the use of two subunits (S1 and S2) of spike for detection of antibody to SARS-CoV.
Identifieur interne : 000305 ( France/Analysis ); précédent : 000304; suivant : 000306False-positive results in a recombinant severe acute respiratory syndrome-associated coronavirus (SARS-CoV) nucleocapsid-based western blot assay were rectified by the use of two subunits (S1 and S2) of spike for detection of antibody to SARS-CoV.
Auteurs : Mimoun Maache [France] ; Florence Komurian-Pradel ; Alain Rajoharison ; Magali Perret ; Jean-Luc Berland ; Stéphane Pouzol ; Audrey Bagnaud ; Blandine Duverger ; Jianguo Xu ; Antonio Osuna ; Glaucia Paranhos-BaccalàSource :
- Clinical and vaccine immunology : CVI [ 1556-6811 ] ; 2006.
Descripteurs français
- KwdFr :
- Anticorps antiviraux (analyse), Anticorps antiviraux (biosynthèse), Faux positifs, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (immunologie), Humains, Protéines de l'enveloppe virale (immunologie), Protéines nucléocapside (biosynthèse), Protéines nucléocapside (génétique), Protéines nucléocapside (immunologie), Protéines nucléocapside (isolement et purification), Protéines recombinantes (biosynthèse), Protéines recombinantes (génétique), Protéines recombinantes (immunologie), Protéines recombinantes (isolement et purification), Sous-unités de protéines (biosynthèse), Sous-unités de protéines (génétique), Sous-unités de protéines (immunologie), Sous-unités de protéines (isolement et purification), Syndrome respiratoire aigu sévère (immunologie), Syndrome respiratoire aigu sévère (virologie), Technique de Western (), Virus du SRAS (immunologie).
- MESH :
- analyse : Anticorps antiviraux.
- biosynthèse : Anticorps antiviraux, Protéines nucléocapside, Protéines recombinantes, Sous-unités de protéines.
- génétique : Protéines nucléocapside, Protéines recombinantes, Sous-unités de protéines.
- immunologie : Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines nucléocapside, Protéines recombinantes, Sous-unités de protéines, Syndrome respiratoire aigu sévère, Virus du SRAS.
- isolement et purification : Protéines nucléocapside, Protéines recombinantes, Sous-unités de protéines.
- virologie : Syndrome respiratoire aigu sévère.
- Faux positifs, Glycoprotéine de spicule des coronavirus, Humains, Technique de Western.
English descriptors
- KwdEn :
- Antibodies, Viral (analysis), Antibodies, Viral (biosynthesis), Blotting, Western (methods), False Positive Reactions, Humans, Membrane Glycoproteins (immunology), Nucleocapsid Proteins (biosynthesis), Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (immunology), Nucleocapsid Proteins (isolation & purification), Protein Subunits (biosynthesis), Protein Subunits (genetics), Protein Subunits (immunology), Protein Subunits (isolation & purification), Recombinant Proteins (biosynthesis), Recombinant Proteins (genetics), Recombinant Proteins (immunology), Recombinant Proteins (isolation & purification), SARS Virus (immunology), Severe Acute Respiratory Syndrome (immunology), Severe Acute Respiratory Syndrome (virology), Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (immunology).
- MESH :
- chemical , analysis : Antibodies, Viral.
- chemical , biosynthesis : Antibodies, Viral, Nucleocapsid Proteins, Protein Subunits, Recombinant Proteins.
- chemical , genetics : Nucleocapsid Proteins, Protein Subunits, Recombinant Proteins.
- chemical , immunology : Membrane Glycoproteins, Nucleocapsid Proteins, Protein Subunits, Recombinant Proteins, Viral Envelope Proteins.
- chemical , isolation & purification : Nucleocapsid Proteins, Protein Subunits, Recombinant Proteins.
- immunology : SARS Virus, Severe Acute Respiratory Syndrome.
- methods : Blotting, Western.
- virology : Severe Acute Respiratory Syndrome.
- False Positive Reactions, Humans, Spike Glycoprotein, Coronavirus.
Abstract
To evaluate the reactivity of the recombinant proteins expressed in Escherichia coli strain BL21(DE3), a Western blot assay was performed by using a panel of 78 serum samples obtained, respectively, from convalescent-phase patients infected with severe acute respiratory syndrome-associated coronavirus (SARS-CoV) (30 samples) and from healthy donors (48 samples). As antigen for detection of SARS-CoV, the nucleocapsid protein (N) showed high sensitivity and strong reactivity with all samples from SARS-CoV patients and cross-reacted with all serum samples from healthy subjects, with either those obtained from China (10 samples) or those obtained from France (38 serum samples), giving then a significant rate of false positives. Specifically, our data indicated that the two subunits, S1 (residues 14 to 760) and S2 (residues 761 to 1190), resulted from the divided spike reacted with all samples from SARS-CoV patients and without any cross-reactivity with any of the healthy serum samples. Consequently, these data revealed the nonspecific nature of N protein in serodiagnosis of SARS-CoV compared with the S1 and S2, where the specificity is of 100%. Moreover, the reported results indicated that the use of one single protein as a detection antigen of SARS-CoV infection may lead to false-positive diagnosis. These may be rectified by using more than one protein for the serodiagnosis of SARS-CoV.
DOI: 10.1128/CVI.13.3.409-414.2006
PubMed: 16522785
Affiliations:
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pubmed:16522785Le document en format XML
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use of two subunits (S1 and S2) of spike for detection of antibody to SARS-CoV.</title><author><name sortKey="Maache, Mimoun" sort="Maache, Mimoun" uniqKey="Maache M" first="Mimoun" last="Maache">Mimoun Maache</name><affiliation wicri:level="3"><nlm:affiliation>Emerging Pathogens Department of bioMérieux, CERVI, 21 Avenue Tony Garnier, 69365 cedex 07, Lyon, France. mmaache@ugr.es</nlm:affiliation><country xml:lang="fr">France</country><wicri:regionArea>Emerging Pathogens Department of bioMérieux, CERVI, 21 Avenue Tony Garnier, 69365 cedex 07, Lyon</wicri:regionArea><placeName><region type="region">Auvergne-Rhône-Alpes</region><region type="old region">Rhône-Alpes</region><settlement type="city">Lyon</settlement></placeName></affiliation></author><author><name sortKey="Komurian Pradel, Florence" sort="Komurian Pradel, Florence" uniqKey="Komurian Pradel F" first="Florence" last="Komurian-Pradel">Florence Komurian-Pradel</name></author><author><name sortKey="Rajoharison, Alain" sort="Rajoharison, Alain" uniqKey="Rajoharison A" first="Alain" last="Rajoharison">Alain Rajoharison</name></author><author><name sortKey="Perret, Magali" sort="Perret, Magali" uniqKey="Perret M" first="Magali" last="Perret">Magali Perret</name></author><author><name sortKey="Berland, Jean Luc" sort="Berland, Jean Luc" uniqKey="Berland J" first="Jean-Luc" last="Berland">Jean-Luc Berland</name></author><author><name sortKey="Pouzol, Stephane" sort="Pouzol, Stephane" uniqKey="Pouzol S" first="Stéphane" last="Pouzol">Stéphane Pouzol</name></author><author><name sortKey="Bagnaud, Audrey" sort="Bagnaud, Audrey" uniqKey="Bagnaud A" first="Audrey" last="Bagnaud">Audrey Bagnaud</name></author><author><name sortKey="Duverger, Blandine" sort="Duverger, Blandine" uniqKey="Duverger B" first="Blandine" last="Duverger">Blandine Duverger</name></author><author><name sortKey="Xu, Jianguo" sort="Xu, Jianguo" uniqKey="Xu J" first="Jianguo" last="Xu">Jianguo Xu</name></author><author><name sortKey="Osuna, Antonio" sort="Osuna, Antonio" uniqKey="Osuna A" first="Antonio" last="Osuna">Antonio Osuna</name></author><author><name sortKey="Paranhos Baccala, Glaucia" sort="Paranhos Baccala, Glaucia" uniqKey="Paranhos Baccala G" first="Glaucia" last="Paranhos-Baccalà">Glaucia Paranhos-Baccalà</name></author></analytic><series><title level="j">Clinical and vaccine immunology : CVI</title><idno type="ISSN">1556-6811</idno><imprint><date when="2006" type="published">2006</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibodies, Viral (analysis)</term><term>Antibodies, Viral (biosynthesis)</term><term>Blotting, Western (methods)</term><term>False Positive Reactions</term><term>Humans</term><term>Membrane Glycoproteins (immunology)</term><term>Nucleocapsid Proteins (biosynthesis)</term><term>Nucleocapsid Proteins (genetics)</term><term>Nucleocapsid Proteins (immunology)</term><term>Nucleocapsid 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scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Antibodies, Viral</term><term>Nucleocapsid Proteins</term><term>Protein Subunits</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Nucleocapsid Proteins</term><term>Protein Subunits</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Membrane Glycoproteins</term><term>Nucleocapsid Proteins</term><term>Protein Subunits</term><term>Recombinant Proteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Nucleocapsid Proteins</term><term>Protein Subunits</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Anticorps antiviraux</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" 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protéines</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Blotting, Western</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>False Positive Reactions</term><term>Humans</term><term>Spike Glycoprotein, Coronavirus</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Faux positifs</term><term>Glycoprotéine de spicule des coronavirus</term><term>Humains</term><term>Technique de Western</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">To evaluate the reactivity of the recombinant proteins expressed in Escherichia coli strain BL21(DE3), a Western blot assay was performed by using a panel of 78 serum samples obtained, respectively, from convalescent-phase patients infected with severe acute respiratory syndrome-associated coronavirus (SARS-CoV) (30 samples) and from healthy donors (48 samples). As antigen for detection of SARS-CoV, the nucleocapsid protein (N) showed high sensitivity and strong reactivity with all samples from SARS-CoV patients and cross-reacted with all serum samples from healthy subjects, with either those obtained from China (10 samples) or those obtained from France (38 serum samples), giving then a significant rate of false positives. Specifically, our data indicated that the two subunits, S1 (residues 14 to 760) and S2 (residues 761 to 1190), resulted from the divided spike reacted with all samples from SARS-CoV patients and without any cross-reactivity with any of the healthy serum samples. Consequently, these data revealed the nonspecific nature of N protein in serodiagnosis of SARS-CoV compared with the S1 and S2, where the specificity is of 100%. Moreover, the reported results indicated that the use of one single protein as a detection antigen of SARS-CoV infection may lead to false-positive diagnosis. These may be rectified by using more than one protein for the serodiagnosis of SARS-CoV.</div></front></TEI><affiliations><list><country><li>France</li></country><region><li>Auvergne-Rhône-Alpes</li><li>Rhône-Alpes</li></region><settlement><li>Lyon</li></settlement></list><tree><noCountry><name sortKey="Bagnaud, Audrey" sort="Bagnaud, Audrey" uniqKey="Bagnaud A" first="Audrey" last="Bagnaud">Audrey Bagnaud</name><name sortKey="Berland, Jean Luc" sort="Berland, Jean Luc" uniqKey="Berland J" first="Jean-Luc" last="Berland">Jean-Luc Berland</name><name sortKey="Duverger, Blandine" sort="Duverger, Blandine" uniqKey="Duverger B" first="Blandine" last="Duverger">Blandine Duverger</name><name sortKey="Komurian Pradel, Florence" sort="Komurian Pradel, Florence" uniqKey="Komurian Pradel F" first="Florence" last="Komurian-Pradel">Florence Komurian-Pradel</name><name sortKey="Osuna, Antonio" sort="Osuna, Antonio" uniqKey="Osuna A" first="Antonio" last="Osuna">Antonio Osuna</name><name sortKey="Paranhos Baccala, Glaucia" sort="Paranhos Baccala, Glaucia" uniqKey="Paranhos Baccala G" first="Glaucia" last="Paranhos-Baccalà">Glaucia Paranhos-Baccalà</name><name sortKey="Perret, Magali" sort="Perret, Magali" uniqKey="Perret M" first="Magali" last="Perret">Magali Perret</name><name sortKey="Pouzol, Stephane" sort="Pouzol, Stephane" uniqKey="Pouzol S" first="Stéphane" last="Pouzol">Stéphane Pouzol</name><name sortKey="Rajoharison, Alain" sort="Rajoharison, Alain" uniqKey="Rajoharison A" first="Alain" last="Rajoharison">Alain Rajoharison</name><name sortKey="Xu, Jianguo" sort="Xu, Jianguo" uniqKey="Xu J" first="Jianguo" last="Xu">Jianguo Xu</name></noCountry><country name="France"><region name="Auvergne-Rhône-Alpes"><name sortKey="Maache, Mimoun" sort="Maache, Mimoun" uniqKey="Maache M" first="Mimoun" last="Maache">Mimoun Maache</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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