Sensitive and specific enzyme-linked immunosorbent assay using chemiluminescence for detection of severe acute respiratory syndrome viral infection.
Identifieur interne : 001C73 ( PubMed/Corpus ); précédent : 001C72; suivant : 001C74Sensitive and specific enzyme-linked immunosorbent assay using chemiluminescence for detection of severe acute respiratory syndrome viral infection.
Auteurs : Kotaro Fujimoto ; Kwok-Hung Chan ; Kazuhiko Takeda ; Kam-Fai Lo ; Raymond H K. Leung ; Takashi OkamotoSource :
- Journal of clinical microbiology [ 1098-660X ] ; 2008.
English descriptors
- KwdEn :
- Adolescent, Adult, Aged, Antibodies, Monoclonal (isolation & purification), Antibodies, Viral (isolation & purification), Antigens, Viral (analysis), Enzyme-Linked Immunosorbent Assay (methods), Female, Humans, Luminescence, Male, Middle Aged, Nucleocapsid Proteins (analysis), Pharynx (virology), Polymerase Chain Reaction (methods), SARS Virus (chemistry), SARS Virus (isolation & purification), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis).
- MESH :
- chemical , analysis : Antigens, Viral, Nucleocapsid Proteins.
- chemical , isolation & purification : Antibodies, Monoclonal, Antibodies, Viral.
- chemistry : SARS Virus.
- diagnosis : Severe Acute Respiratory Syndrome.
- isolation & purification : SARS Virus.
- methods : Enzyme-Linked Immunosorbent Assay, Polymerase Chain Reaction.
- virology : Pharynx.
- Adolescent, Adult, Aged, Female, Humans, Luminescence, Male, Middle Aged, Sensitivity and Specificity.
Abstract
Here we report the development of a more-sensitive immunoassay for severe acute respiratory syndrome (SARS) based on an enzyme-linked immunosorbent assay using chemiluminescence (CLEIA) to detect the viral nucleocapsid (N) antigen in nasopharyngeal aspirate (NPA) from patients infected with SARS coronavirus (CoV). The CLEIA was established with an optical combination of monoclonal antibodies (MAbs) against SARS CoV N protein prepared from mice immunized with recombinant N protein without cultivating the virus. The capture and detecting MAbs of the CLEIA reacted to the carboxyl-terminal and amino-terminal peptides of the N protein, respectively. The CLEIA was capable of detecting recombinant N protein at 1.56 pg/ml and viral N protein in SARS CoV cell culture lysates at 0.087 of 50% tissue culture infective doses/ml. The CLEIA showed no cross-reactivities to recombinant N proteins of common human CoV (229E, OC43, and NL63) or lysates of cells infected with 229E and OC43. In addition, an evaluation with 18 SARS-positive NPA samples, all confirmed SARS positive by quantitative PCR and antibodies to SARS CoV, revealed that all (18/18) were found positive by the CLEIA; thus, the sensitivity of detection was 100%. When we tested 20 SARS-negative NPA samples, the CLEIA was shown to have high specificity (100%). The sensitivity of our novel SARS CLEIA was significantly higher than the previous EIA and comparable to the other methods using reverse transcription-PCR.
DOI: 10.1128/JCM.01006-07
PubMed: 18032623
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pubmed:18032623Le document en format XML
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Ltd., Kobe, Japan.</nlm:affiliation></affiliation></author><author><name sortKey="Chan, Kwok Hung" sort="Chan, Kwok Hung" uniqKey="Chan K" first="Kwok-Hung" last="Chan">Kwok-Hung Chan</name></author><author><name sortKey="Takeda, Kazuhiko" sort="Takeda, Kazuhiko" uniqKey="Takeda K" first="Kazuhiko" last="Takeda">Kazuhiko Takeda</name></author><author><name sortKey="Lo, Kam Fai" sort="Lo, Kam Fai" uniqKey="Lo K" first="Kam-Fai" last="Lo">Kam-Fai Lo</name></author><author><name sortKey="Leung, Raymond H K" sort="Leung, Raymond H K" uniqKey="Leung R" first="Raymond H K" last="Leung">Raymond H K. Leung</name></author><author><name sortKey="Okamoto, Takashi" sort="Okamoto, Takashi" uniqKey="Okamoto T" first="Takashi" last="Okamoto">Takashi Okamoto</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2008">2008</date><idno type="RBID">pubmed:18032623</idno><idno type="pmid">18032623</idno><idno type="doi">10.1128/JCM.01006-07</idno><idno type="wicri:Area/PubMed/Corpus">001C73</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001C73</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Sensitive and specific enzyme-linked immunosorbent assay using chemiluminescence for detection of severe acute respiratory syndrome viral infection.</title><author><name sortKey="Fujimoto, Kotaro" sort="Fujimoto, Kotaro" uniqKey="Fujimoto K" first="Kotaro" last="Fujimoto">Kotaro Fujimoto</name><affiliation><nlm:affiliation>Research and Development Division, Sysmex Co. Ltd., Kobe, Japan.</nlm:affiliation></affiliation></author><author><name sortKey="Chan, Kwok Hung" sort="Chan, Kwok Hung" uniqKey="Chan K" first="Kwok-Hung" last="Chan">Kwok-Hung Chan</name></author><author><name sortKey="Takeda, Kazuhiko" sort="Takeda, Kazuhiko" uniqKey="Takeda K" first="Kazuhiko" last="Takeda">Kazuhiko Takeda</name></author><author><name sortKey="Lo, Kam Fai" sort="Lo, Kam Fai" uniqKey="Lo K" first="Kam-Fai" last="Lo">Kam-Fai Lo</name></author><author><name sortKey="Leung, Raymond H K" sort="Leung, Raymond H K" uniqKey="Leung R" first="Raymond H K" last="Leung">Raymond H K. Leung</name></author><author><name sortKey="Okamoto, Takashi" sort="Okamoto, Takashi" uniqKey="Okamoto T" first="Takashi" last="Okamoto">Takashi Okamoto</name></author></analytic><series><title level="j">Journal of clinical microbiology</title><idno type="eISSN">1098-660X</idno><imprint><date when="2008" type="published">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Adolescent</term><term>Adult</term><term>Aged</term><term>Antibodies, Monoclonal (isolation & purification)</term><term>Antibodies, Viral (isolation & purification)</term><term>Antigens, Viral (analysis)</term><term>Enzyme-Linked Immunosorbent Assay (methods)</term><term>Female</term><term>Humans</term><term>Luminescence</term><term>Male</term><term>Middle Aged</term><term>Nucleocapsid Proteins (analysis)</term><term>Pharynx (virology)</term><term>Polymerase Chain Reaction (methods)</term><term>SARS Virus (chemistry)</term><term>SARS Virus (isolation & purification)</term><term>Sensitivity and Specificity</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Antigens, Viral</term><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Antibodies, Monoclonal</term><term>Antibodies, Viral</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Enzyme-Linked Immunosorbent Assay</term><term>Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Pharynx</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Adolescent</term><term>Adult</term><term>Aged</term><term>Female</term><term>Humans</term><term>Luminescence</term><term>Male</term><term>Middle Aged</term><term>Sensitivity and Specificity</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Here we report the development of a more-sensitive immunoassay for severe acute respiratory syndrome (SARS) based on an enzyme-linked immunosorbent assay using chemiluminescence (CLEIA) to detect the viral nucleocapsid (N) antigen in nasopharyngeal aspirate (NPA) from patients infected with SARS coronavirus (CoV). The CLEIA was established with an optical combination of monoclonal antibodies (MAbs) against SARS CoV N protein prepared from mice immunized with recombinant N protein without cultivating the virus. The capture and detecting MAbs of the CLEIA reacted to the carboxyl-terminal and amino-terminal peptides of the N protein, respectively. The CLEIA was capable of detecting recombinant N protein at 1.56 pg/ml and viral N protein in SARS CoV cell culture lysates at 0.087 of 50% tissue culture infective doses/ml. The CLEIA showed no cross-reactivities to recombinant N proteins of common human CoV (229E, OC43, and NL63) or lysates of cells infected with 229E and OC43. In addition, an evaluation with 18 SARS-positive NPA samples, all confirmed SARS positive by quantitative PCR and antibodies to SARS CoV, revealed that all (18/18) were found positive by the CLEIA; thus, the sensitivity of detection was 100%. When we tested 20 SARS-negative NPA samples, the CLEIA was shown to have high specificity (100%). The sensitivity of our novel SARS CLEIA was significantly higher than the previous EIA and comparable to the other methods using reverse transcription-PCR.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">18032623</PMID><DateCompleted><Year>2008</Year><Month>04</Month><Day>08</Day></DateCompleted><DateRevised><Year>2019</Year><Month>12</Month><Day>10</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1098-660X</ISSN><JournalIssue CitedMedium="Internet"><Volume>46</Volume><Issue>1</Issue><PubDate><Year>2008</Year><Month>Jan</Month></PubDate></JournalIssue><Title>Journal of clinical microbiology</Title><ISOAbbreviation>J. Clin. Microbiol.</ISOAbbreviation></Journal><ArticleTitle>Sensitive and specific enzyme-linked immunosorbent assay using chemiluminescence for detection of severe acute respiratory syndrome viral infection.</ArticleTitle><Pagination><MedlinePgn>302-10</MedlinePgn></Pagination><Abstract><AbstractText>Here we report the development of a more-sensitive immunoassay for severe acute respiratory syndrome (SARS) based on an enzyme-linked immunosorbent assay using chemiluminescence (CLEIA) to detect the viral nucleocapsid (N) antigen in nasopharyngeal aspirate (NPA) from patients infected with SARS coronavirus (CoV). The CLEIA was established with an optical combination of monoclonal antibodies (MAbs) against SARS CoV N protein prepared from mice immunized with recombinant N protein without cultivating the virus. The capture and detecting MAbs of the CLEIA reacted to the carboxyl-terminal and amino-terminal peptides of the N protein, respectively. The CLEIA was capable of detecting recombinant N protein at 1.56 pg/ml and viral N protein in SARS CoV cell culture lysates at 0.087 of 50% tissue culture infective doses/ml. The CLEIA showed no cross-reactivities to recombinant N proteins of common human CoV (229E, OC43, and NL63) or lysates of cells infected with 229E and OC43. In addition, an evaluation with 18 SARS-positive NPA samples, all confirmed SARS positive by quantitative PCR and antibodies to SARS CoV, revealed that all (18/18) were found positive by the CLEIA; thus, the sensitivity of detection was 100%. When we tested 20 SARS-negative NPA samples, the CLEIA was shown to have high specificity (100%). The sensitivity of our novel SARS CLEIA was significantly higher than the previous EIA and comparable to the other methods using reverse transcription-PCR.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Fujimoto</LastName><ForeName>Kotaro</ForeName><Initials>K</Initials><AffiliationInfo><Affiliation>Research and Development Division, Sysmex Co. Ltd., Kobe, Japan.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Chan</LastName><ForeName>Kwok-Hung</ForeName><Initials>KH</Initials></Author><Author ValidYN="Y"><LastName>Takeda</LastName><ForeName>Kazuhiko</ForeName><Initials>K</Initials></Author><Author ValidYN="Y"><LastName>Lo</LastName><ForeName>Kam-Fai</ForeName><Initials>KF</Initials></Author><Author ValidYN="Y"><LastName>Leung</LastName><ForeName>Raymond H K</ForeName><Initials>RH</Initials></Author><Author ValidYN="Y"><LastName>Okamoto</LastName><ForeName>Takashi</ForeName><Initials>T</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D003160">Comparative Study</PublicationType><PublicationType UI="D023362">Evaluation Study</PublicationType><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate 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