Development of an enzyme-linked immunosorbent assay-based test with a cocktail of nucleocapsid and spike proteins for detection of severe acute respiratory syndrome-associated coronavirus-specific antibody.
Identifieur interne : 001A24 ( PubMed/Corpus ); précédent : 001A23; suivant : 001A25Development of an enzyme-linked immunosorbent assay-based test with a cocktail of nucleocapsid and spike proteins for detection of severe acute respiratory syndrome-associated coronavirus-specific antibody.
Auteurs : Luis G. Giménez ; Jose Rojas ; Almudena Rojas ; Joaquín Mendoza ; Ana G. CamachoSource :
- Clinical and vaccine immunology : CVI [ 1556-679X ] ; 2009.
English descriptors
- KwdEn :
- Antibodies, Viral (blood), Antigens, Viral, Enzyme-Linked Immunosorbent Assay (methods), Humans, Immunoglobulin G (blood), Immunoglobulin M (blood), Membrane Glycoproteins (immunology), Nucleocapsid Proteins (immunology), Recombinant Proteins, SARS Virus (immunology), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (immunology).
- MESH :
- chemical , blood : Antibodies, Viral, Immunoglobulin G, Immunoglobulin M.
- chemical , immunology : Membrane Glycoproteins, Nucleocapsid Proteins, Viral Envelope Proteins.
- chemical : Antigens, Viral, Recombinant Proteins, Spike Glycoprotein, Coronavirus.
- diagnosis : Severe Acute Respiratory Syndrome.
- immunology : SARS Virus.
- methods : Enzyme-Linked Immunosorbent Assay.
- Humans, Sensitivity and Specificity.
Abstract
A new enzyme-linked immunosorbent assay (ELISA)-based immunoglobulin G (IgG)-plus-IgM antibody detection test for severe acute respiratory syndrome (SARS) has been developed by using a cocktail of four recombinant polypeptides as the antigen. These recombinant fragments were designed as parts of two different structural proteins from SARS-associated coronavirus (SARS-CoV). One recombinant polypeptide, S251-683, was designed as part of the spike glycoprotein, and the other three polypeptides comprised almost the whole nucleocapsid protein, avoiding the last 25 C-terminal amino acids. Immunization with a cocktail of these four polypeptides yielded a specific polyclonal antibody that is able to recognize SARS-CoV-infected cells by an immunofluorescence assay. This polypeptide cocktail was also used to set up an ELISA-based IgG-plus-IgM antibody detection test, which showed 99% specificity and 90% sensitivity upon evaluation using sera from 100 healthy negative controls and 20 SARS patients. Separate immunoreactivity assays with each recombinant polypeptide demonstrated that a combination of N and S protein fragments was more suitable than the individual peptides for developing a serological assay for SARS-CoV.
DOI: 10.1128/CVI.00252-08
PubMed: 19038782
Links to Exploration step
pubmed:19038782Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Development of an enzyme-linked immunosorbent assay-based test with a cocktail of nucleocapsid and spike proteins for detection of severe acute respiratory syndrome-associated coronavirus-specific antibody.</title><author><name sortKey="Gimenez, Luis G" sort="Gimenez, Luis G" uniqKey="Gimenez L" first="Luis G" last="Giménez">Luis G. Giménez</name><affiliation><nlm:affiliation>Laboratorios Vircell, SL, Granada, Spain.</nlm:affiliation></affiliation></author><author><name sortKey="Rojas, Jose" sort="Rojas, Jose" uniqKey="Rojas J" first="Jose" last="Rojas">Jose Rojas</name></author><author><name sortKey="Rojas, Almudena" sort="Rojas, Almudena" uniqKey="Rojas A" first="Almudena" last="Rojas">Almudena Rojas</name></author><author><name sortKey="Mendoza, Joaquin" sort="Mendoza, Joaquin" uniqKey="Mendoza J" first="Joaquín" last="Mendoza">Joaquín Mendoza</name></author><author><name sortKey="Camacho, Ana G" sort="Camacho, Ana G" uniqKey="Camacho A" first="Ana G" last="Camacho">Ana G. Camacho</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2009">2009</date><idno type="RBID">pubmed:19038782</idno><idno type="pmid">19038782</idno><idno type="doi">10.1128/CVI.00252-08</idno><idno type="wicri:Area/PubMed/Corpus">001A24</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001A24</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Development of an enzyme-linked immunosorbent assay-based test with a cocktail of nucleocapsid and spike proteins for detection of severe acute respiratory syndrome-associated coronavirus-specific antibody.</title><author><name sortKey="Gimenez, Luis G" sort="Gimenez, Luis G" uniqKey="Gimenez L" first="Luis G" last="Giménez">Luis G. Giménez</name><affiliation><nlm:affiliation>Laboratorios Vircell, SL, Granada, Spain.</nlm:affiliation></affiliation></author><author><name sortKey="Rojas, Jose" sort="Rojas, Jose" uniqKey="Rojas J" first="Jose" last="Rojas">Jose Rojas</name></author><author><name sortKey="Rojas, Almudena" sort="Rojas, Almudena" uniqKey="Rojas A" first="Almudena" last="Rojas">Almudena Rojas</name></author><author><name sortKey="Mendoza, Joaquin" sort="Mendoza, Joaquin" uniqKey="Mendoza J" first="Joaquín" last="Mendoza">Joaquín Mendoza</name></author><author><name sortKey="Camacho, Ana G" sort="Camacho, Ana G" uniqKey="Camacho A" first="Ana G" last="Camacho">Ana G. Camacho</name></author></analytic><series><title level="j">Clinical and vaccine immunology : CVI</title><idno type="eISSN">1556-679X</idno><imprint><date when="2009" type="published">2009</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibodies, Viral (blood)</term><term>Antigens, Viral</term><term>Enzyme-Linked Immunosorbent Assay (methods)</term><term>Humans</term><term>Immunoglobulin G (blood)</term><term>Immunoglobulin M (blood)</term><term>Membrane Glycoproteins (immunology)</term><term>Nucleocapsid Proteins (immunology)</term><term>Recombinant Proteins</term><term>SARS Virus (immunology)</term><term>Sensitivity and Specificity</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term><term>Spike Glycoprotein, Coronavirus</term><term>Viral Envelope Proteins (immunology)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>Antibodies, Viral</term><term>Immunoglobulin G</term><term>Immunoglobulin M</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Membrane Glycoproteins</term><term>Nucleocapsid Proteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Antigens, Viral</term><term>Recombinant Proteins</term><term>Spike Glycoprotein, Coronavirus</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Enzyme-Linked Immunosorbent Assay</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Humans</term><term>Sensitivity and Specificity</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A new enzyme-linked immunosorbent assay (ELISA)-based immunoglobulin G (IgG)-plus-IgM antibody detection test for severe acute respiratory syndrome (SARS) has been developed by using a cocktail of four recombinant polypeptides as the antigen. These recombinant fragments were designed as parts of two different structural proteins from SARS-associated coronavirus (SARS-CoV). One recombinant polypeptide, S251-683, was designed as part of the spike glycoprotein, and the other three polypeptides comprised almost the whole nucleocapsid protein, avoiding the last 25 C-terminal amino acids. Immunization with a cocktail of these four polypeptides yielded a specific polyclonal antibody that is able to recognize SARS-CoV-infected cells by an immunofluorescence assay. This polypeptide cocktail was also used to set up an ELISA-based IgG-plus-IgM antibody detection test, which showed 99% specificity and 90% sensitivity upon evaluation using sera from 100 healthy negative controls and 20 SARS patients. Separate immunoreactivity assays with each recombinant polypeptide demonstrated that a combination of N and S protein fragments was more suitable than the individual peptides for developing a serological assay for SARS-CoV.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">19038782</PMID><DateCompleted><Year>2009</Year><Month>03</Month><Day>09</Day></DateCompleted><DateRevised><Year>2020</Year><Month>04</Month><Day>15</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1556-679X</ISSN><JournalIssue CitedMedium="Internet"><Volume>16</Volume><Issue>2</Issue><PubDate><Year>2009</Year><Month>Feb</Month></PubDate></JournalIssue><Title>Clinical and vaccine immunology : CVI</Title><ISOAbbreviation>Clin. Vaccine Immunol.</ISOAbbreviation></Journal><ArticleTitle>Development of an enzyme-linked immunosorbent assay-based test with a cocktail of nucleocapsid and spike proteins for detection of severe acute respiratory syndrome-associated coronavirus-specific antibody.</ArticleTitle><Pagination><MedlinePgn>241-5</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1128/CVI.00252-08</ELocationID><Abstract><AbstractText>A new enzyme-linked immunosorbent assay (ELISA)-based immunoglobulin G (IgG)-plus-IgM antibody detection test for severe acute respiratory syndrome (SARS) has been developed by using a cocktail of four recombinant polypeptides as the antigen. These recombinant fragments were designed as parts of two different structural proteins from SARS-associated coronavirus (SARS-CoV). One recombinant polypeptide, S251-683, was designed as part of the spike glycoprotein, and the other three polypeptides comprised almost the whole nucleocapsid protein, avoiding the last 25 C-terminal amino acids. Immunization with a cocktail of these four polypeptides yielded a specific polyclonal antibody that is able to recognize SARS-CoV-infected cells by an immunofluorescence assay. This polypeptide cocktail was also used to set up an ELISA-based IgG-plus-IgM antibody detection test, which showed 99% specificity and 90% sensitivity upon evaluation using sera from 100 healthy negative controls and 20 SARS patients. Separate immunoreactivity assays with each recombinant polypeptide demonstrated that a combination of N and S protein fragments was more suitable than the individual peptides for developing a serological assay for SARS-CoV.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Giménez</LastName><ForeName>Luis G</ForeName><Initials>LG</Initials><AffiliationInfo><Affiliation>Laboratorios Vircell, SL, Granada, Spain.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Rojas</LastName><ForeName>Jose</ForeName><Initials>J</Initials></Author><Author ValidYN="Y"><LastName>Rojas</LastName><ForeName>Almudena</ForeName><Initials>A</Initials></Author><Author ValidYN="Y"><LastName>Mendoza</LastName><ForeName>Joaquín</ForeName><Initials>J</Initials></Author><Author ValidYN="Y"><LastName>Camacho</LastName><ForeName>Ana G</ForeName><Initials>AG</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D023362">Evaluation Study</PublicationType><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2008</Year><Month>11</Month><Day>26</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Clin Vaccine Immunol</MedlineTA><NlmUniqueID>101252125</NlmUniqueID><ISSNLinking>1556-679X</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="D007075">Immunoglobulin M</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C578553">MHV 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