Screening of specific antigens for SARS clinical diagnosis using a protein microarray.
Identifieur interne : 000E44 ( Ncbi/Merge ); précédent : 000E43; suivant : 000E45Screening of specific antigens for SARS clinical diagnosis using a protein microarray.
Auteurs : Dan-Dan Lu [République populaire de Chine] ; Su-Hong Chen ; Shi-Meng Zhang ; Min-Li Zhang ; Wei Zhang ; Xiao-Chen Bo ; Sheng-Qi WangSource :
- The Analyst [ 0003-2654 ] ; 2005.
Descripteurs français
- KwdFr :
- Analyse par réseau de protéines, Anticorps antiviraux (analyse), Antigènes viraux (isolement et purification), Bioréacteurs, Escherichia coli, Humains, Immunoglobuline G (analyse), Protéines de fusion recombinantes (analyse), Protéines et peptides de signalisation intracellulaire (génétique), Protéines membranaires, Sensibilité et spécificité, Spectrométrie de masse MALDI, Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (virologie), Test ELISA (), Transporteurs d'anions organiques (génétique), Virus du SRAS (immunologie), Électrophorèse sur gel de polyacrylamide, Études cas-témoins.
- MESH :
- analyse : Anticorps antiviraux, Immunoglobuline G, Protéines de fusion recombinantes.
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Protéines et peptides de signalisation intracellulaire, Transporteurs d'anions organiques.
- immunologie : Virus du SRAS.
- isolement et purification : Antigènes viraux.
- virologie : Syndrome respiratoire aigu sévère.
- Analyse par réseau de protéines, Bioréacteurs, Escherichia coli, Humains, Protéines membranaires, Sensibilité et spécificité, Spectrométrie de masse MALDI, Test ELISA, Électrophorèse sur gel de polyacrylamide, Études cas-témoins.
English descriptors
- KwdEn :
- Antibodies, Viral (analysis), Antigens, Viral (isolation & purification), Bioreactors, Case-Control Studies, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay (methods), Escherichia coli, Humans, Immunoglobulin G (analysis), Intracellular Signaling Peptides and Proteins (genetics), Membrane Proteins, Organic Anion Transporters (genetics), Protein Array Analysis, Recombinant Fusion Proteins (analysis), SARS Virus (immunology), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (virology), Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization.
- MESH :
- chemical , analysis : Antibodies, Viral, Immunoglobulin G, Recombinant Fusion Proteins.
- chemical , genetics : Intracellular Signaling Peptides and Proteins, Organic Anion Transporters.
- chemical , isolation & purification : Antigens, Viral.
- diagnosis : Severe Acute Respiratory Syndrome.
- immunology : SARS Virus.
- methods : Enzyme-Linked Immunosorbent Assay.
- virology : Severe Acute Respiratory Syndrome.
- Bioreactors, Case-Control Studies, Electrophoresis, Polyacrylamide Gel, Escherichia coli, Humans, Membrane Proteins, Protein Array Analysis, Sensitivity and Specificity, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization.
Abstract
In this study several SARS-CoV structural proteins and fragments were expressed in E. coli as GST or TRX fusion proteins. They were fabricated on a microarray and tested with sera from SARS patients. Antigenic screening indicated that recombinant GST-N2 fusion protein, the carboxy-terminus 213aa-423aa of N protein, was strongest positive and weakest non-specific compared with others. An indirect antibody ELISA method was developed and clinical positive and negative sera for their antibodies against GST-N2 fusion protein were assayed. 311 out of the 442 sera from clinical SARS inpatients, as well as 229 out of 302 sera from convalescent patients gave positive reactivities; positive rates were 70.4% and 75.8% respectively. Sera from a total of 2726 non-SARS patients and healthy individuals were tested and the false positive rate was only 0.07%. When the sensitivity control sample was diluted 1 : 64, it yielded OD values above the cutoff value. Reported data showed that this was a relatively high degree of sensitivity and specificity for SARS-CoV antibody testing. The data indicate that GST-N2 fusion protein, which was screened by protein microarray, may be a valuable diagnostic antigen for the development of serological assays for SARS. In addition, protein microarray assay presents a higher positive rate and sensitivity (86.1% and 1 : 200) compared with the traditional ELISA screening method, and could provide a rapid, parallel and high-throughput antigen screening platform.
DOI: 10.1039/b415888a
PubMed: 15776156
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Meng" sort="Zhang, Shi Meng" uniqKey="Zhang S" first="Shi-Meng" last="Zhang">Shi-Meng Zhang</name></author><author><name sortKey="Zhang, Min Li" sort="Zhang, Min Li" uniqKey="Zhang M" first="Min-Li" last="Zhang">Min-Li Zhang</name></author><author><name sortKey="Zhang, Wei" sort="Zhang, Wei" uniqKey="Zhang W" first="Wei" last="Zhang">Wei Zhang</name></author><author><name sortKey="Bo, Xiao Chen" sort="Bo, Xiao Chen" uniqKey="Bo X" first="Xiao-Chen" last="Bo">Xiao-Chen Bo</name></author><author><name sortKey="Wang, Sheng Qi" sort="Wang, Sheng Qi" uniqKey="Wang S" first="Sheng-Qi" last="Wang">Sheng-Qi Wang</name></author></analytic><series><title level="j">The Analyst</title><idno type="ISSN">0003-2654</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibodies, Viral (analysis)</term><term>Antigens, Viral (isolation & purification)</term><term>Bioreactors</term><term>Case-Control Studies</term><term>Electrophoresis, Polyacrylamide Gel</term><term>Enzyme-Linked Immunosorbent Assay (methods)</term><term>Escherichia coli</term><term>Humans</term><term>Immunoglobulin G (analysis)</term><term>Intracellular Signaling Peptides and Proteins (genetics)</term><term>Membrane Proteins</term><term>Organic Anion Transporters (genetics)</term><term>Protein Array Analysis</term><term>Recombinant Fusion Proteins (analysis)</term><term>SARS Virus (immunology)</term><term>Sensitivity and Specificity</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term><term>Severe Acute Respiratory Syndrome (virology)</term><term>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Analyse par réseau de protéines</term><term>Anticorps antiviraux (analyse)</term><term>Antigènes viraux (isolement et purification)</term><term>Bioréacteurs</term><term>Escherichia coli</term><term>Humains</term><term>Immunoglobuline G (analyse)</term><term>Protéines de fusion recombinantes (analyse)</term><term>Protéines et peptides de signalisation intracellulaire (génétique)</term><term>Protéines membranaires</term><term>Sensibilité et spécificité</term><term>Spectrométrie de masse MALDI</term><term>Syndrome respiratoire aigu sévère (diagnostic)</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Test ELISA ()</term><term>Transporteurs d'anions organiques (génétique)</term><term>Virus du SRAS (immunologie)</term><term>Électrophorèse sur gel de polyacrylamide</term><term>Études cas-témoins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Antibodies, Viral</term><term>Immunoglobulin G</term><term>Recombinant Fusion Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Intracellular Signaling Peptides and Proteins</term><term>Organic Anion Transporters</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Antigens, Viral</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Anticorps antiviraux</term><term>Immunoglobuline G</term><term>Protéines de fusion recombinantes</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines et peptides de signalisation intracellulaire</term><term>Transporteurs d'anions organiques</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Antigènes viraux</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Enzyme-Linked Immunosorbent Assay</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>Bioreactors</term><term>Case-Control Studies</term><term>Electrophoresis, Polyacrylamide Gel</term><term>Escherichia coli</term><term>Humans</term><term>Membrane Proteins</term><term>Protein Array Analysis</term><term>Sensitivity and Specificity</term><term>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Analyse par réseau de protéines</term><term>Bioréacteurs</term><term>Escherichia coli</term><term>Humains</term><term>Protéines membranaires</term><term>Sensibilité et spécificité</term><term>Spectrométrie de masse MALDI</term><term>Test ELISA</term><term>Électrophorèse sur gel de polyacrylamide</term><term>Études cas-témoins</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">In this study several SARS-CoV structural proteins and fragments were expressed in E. coli as GST or TRX fusion proteins. They were fabricated on a microarray and tested with sera from SARS patients. Antigenic screening indicated that recombinant GST-N2 fusion protein, the carboxy-terminus 213aa-423aa of N protein, was strongest positive and weakest non-specific compared with others. An indirect antibody ELISA method was developed and clinical positive and negative sera for their antibodies against GST-N2 fusion protein were assayed. 311 out of the 442 sera from clinical SARS inpatients, as well as 229 out of 302 sera from convalescent patients gave positive reactivities; positive rates were 70.4% and 75.8% respectively. Sera from a total of 2726 non-SARS patients and healthy individuals were tested and the false positive rate was only 0.07%. When the sensitivity control sample was diluted 1 : 64, it yielded OD values above the cutoff value. Reported data showed that this was a relatively high degree of sensitivity and specificity for SARS-CoV antibody testing. The data indicate that GST-N2 fusion protein, which was screened by protein microarray, may be a valuable diagnostic antigen for the development of serological assays for SARS. In addition, protein microarray assay presents a higher positive rate and sensitivity (86.1% and 1 : 200) compared with the traditional ELISA screening method, and could provide a rapid, parallel and high-throughput antigen screening platform.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">15776156</PMID><DateCompleted><Year>2005</Year><Month>08</Month><Day>11</Day></DateCompleted><DateRevised><Year>2015</Year><Month>11</Month><Day>19</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Print">0003-2654</ISSN><JournalIssue CitedMedium="Print"><Volume>130</Volume><Issue>4</Issue><PubDate><Year>2005</Year><Month>Apr</Month></PubDate></JournalIssue><Title>The Analyst</Title><ISOAbbreviation>Analyst</ISOAbbreviation></Journal><ArticleTitle>Screening of specific antigens for SARS clinical diagnosis using a protein microarray.</ArticleTitle><Pagination><MedlinePgn>474-82</MedlinePgn></Pagination><Abstract><AbstractText>In this study several SARS-CoV structural proteins and fragments were expressed in E. coli as GST or TRX fusion proteins. They were fabricated on a microarray and tested with sera from SARS patients. Antigenic screening indicated that recombinant GST-N2 fusion protein, the carboxy-terminus 213aa-423aa of N protein, was strongest positive and weakest non-specific compared with others. An indirect antibody ELISA method was developed and clinical positive and negative sera for their antibodies against GST-N2 fusion protein were assayed. 311 out of the 442 sera from clinical SARS inpatients, as well as 229 out of 302 sera from convalescent patients gave positive reactivities; positive rates were 70.4% and 75.8% respectively. Sera from a total of 2726 non-SARS patients and healthy individuals were tested and the false positive rate was only 0.07%. When the sensitivity control sample was diluted 1 : 64, it yielded OD values above the cutoff value. Reported data showed that this was a relatively high degree of sensitivity and specificity for SARS-CoV antibody testing. The data indicate that GST-N2 fusion protein, which was screened by protein microarray, may be a valuable diagnostic antigen for the development of serological assays for SARS. In addition, protein microarray assay presents a higher positive rate and sensitivity (86.1% and 1 : 200) compared with the traditional ELISA screening method, and could provide a rapid, parallel and high-throughput antigen screening platform.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Lu</LastName><ForeName>Dan-Dan</ForeName><Initials>DD</Initials><AffiliationInfo><Affiliation>Beijing Institute of Radiation Medicine, Beijing, 100850, People's Republic of China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Chen</LastName><ForeName>Su-Hong</ForeName><Initials>SH</Initials></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Shi-Meng</ForeName><Initials>SM</Initials></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Min-Li</ForeName><Initials>ML</Initials></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Wei</ForeName><Initials>W</Initials></Author><Author ValidYN="Y"><LastName>Bo</LastName><ForeName>Xiao-Chen</ForeName><Initials>XC</Initials></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Sheng-Qi</ForeName><Initials>SQ</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2005</Year><Month>02</Month><Day>14</Day></ArticleDate></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Analyst</MedlineTA><NlmUniqueID>0372652</NlmUniqueID><ISSNLinking>0003-2654</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000914">Antibodies, Viral</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000956">Antigens, Viral</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D007074">Immunoglobulin G</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D047908">Intracellular Signaling Peptides and Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D008565">Membrane Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D027361">Organic Anion Transporters</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011993">Recombinant Fusion Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C494631">VAC14 protein, human</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C487009">gonad-specific transporter, human</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000914" MajorTopicYN="N">Antibodies, Viral</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="Y">analysis</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D000956" MajorTopicYN="N">Antigens, Viral</DescriptorName><QualifierName UI="Q000302" MajorTopicYN="N">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D019149" MajorTopicYN="N">Bioreactors</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D016022" MajorTopicYN="N">Case-Control Studies</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004591" MajorTopicYN="N">Electrophoresis, Polyacrylamide Gel</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004797" MajorTopicYN="N">Enzyme-Linked Immunosorbent Assay</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="N">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004926" MajorTopicYN="N">Escherichia coli</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D007074" MajorTopicYN="N">Immunoglobulin G</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D047908" MajorTopicYN="N">Intracellular Signaling Peptides and Proteins</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D008565" MajorTopicYN="N">Membrane Proteins</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D027361" MajorTopicYN="N">Organic Anion Transporters</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D040081" MajorTopicYN="Y">Protein Array Analysis</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D011993" MajorTopicYN="N">Recombinant Fusion Proteins</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D045473" MajorTopicYN="N">SARS Virus</DescriptorName><QualifierName UI="Q000276" MajorTopicYN="Y">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D012680" MajorTopicYN="N">Sensitivity and Specificity</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D045169" MajorTopicYN="N">Severe Acute Respiratory Syndrome</DescriptorName><QualifierName UI="Q000175" MajorTopicYN="Y">diagnosis</QualifierName><QualifierName UI="Q000821" MajorTopicYN="N">virology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D019032" MajorTopicYN="N">Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2005</Year><Month>3</Month><Day>19</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2005</Year><Month>8</Month><Day>12</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2005</Year><Month>3</Month><Day>19</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">15776156</ArticleId><ArticleId IdType="doi">10.1039/b415888a</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>République populaire de Chine</li></country></list><tree><noCountry><name sortKey="Bo, Xiao Chen" sort="Bo, Xiao Chen" uniqKey="Bo X" first="Xiao-Chen" last="Bo">Xiao-Chen Bo</name><name sortKey="Chen, Su Hong" sort="Chen, Su Hong" uniqKey="Chen S" first="Su-Hong" last="Chen">Su-Hong Chen</name><name sortKey="Wang, Sheng Qi" sort="Wang, Sheng Qi" uniqKey="Wang S" first="Sheng-Qi" last="Wang">Sheng-Qi Wang</name><name sortKey="Zhang, Min Li" sort="Zhang, Min Li" uniqKey="Zhang M" first="Min-Li" last="Zhang">Min-Li Zhang</name><name sortKey="Zhang, Shi Meng" sort="Zhang, Shi Meng" uniqKey="Zhang S" first="Shi-Meng" last="Zhang">Shi-Meng Zhang</name><name sortKey="Zhang, Wei" sort="Zhang, Wei" uniqKey="Zhang W" first="Wei" last="Zhang">Wei Zhang</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Lu, Dan Dan" sort="Lu, Dan Dan" uniqKey="Lu D" first="Dan-Dan" last="Lu">Dan-Dan Lu</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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