SrasV1, Ncbi, Checkpoint, bibRecord, 001B20

Sensitive and Specific Enzyme-Linked Immunosorbent Assay Using Chemiluminescence for Detection of Severe Acute Respiratory Syndrome Viral Infection▿

Identifieur interne : 001B20 ( Ncbi/Checkpoint ); précédent : 001B19; suivant : 001B21

Sensitive 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 Okamoto

Source :

Journal of Clinical Microbiology [ 0095-1137 ] ; 2007.

RBID : PMC:2224272

Descripteurs français

KwdFr :
- Adolescent, Adulte, Adulte d'âge moyen, Anticorps antiviraux (isolement et purification), Anticorps monoclonaux (isolement et purification), Antigènes viraux (analyse), Femelle, Humains, Luminescence, Mâle, Pharynx (virologie), Protéines nucléocapside (analyse), Réaction de polymérisation en chaîne (), Sensibilité et spécificité, Sujet âgé, Syndrome respiratoire aigu sévère (diagnostic), Test ELISA (), Virus du SRAS (), Virus du SRAS (isolement et purification).
MESH :
- analyse : Antigènes viraux, Protéines nucléocapside.
- diagnostic : Syndrome respiratoire aigu sévère.
- isolement et purification : Anticorps antiviraux, Anticorps monoclonaux, Virus du SRAS.
- virologie : Pharynx.
- Adolescent, Adulte, Adulte d'âge moyen, Femelle, Humains, Luminescence, Mâle, Réaction de polymérisation en chaîne, Sensibilité et spécificité, Sujet âgé, Test ELISA, Virus du SRAS.

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.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2224272

DOI: 10.1128/JCM.01006-07
PubMed: 18032623
PubMed Central: 2224272

Affiliations:

Links to Exploration step

PMC:2224272

Le document en format XML

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<term>Antibodies, Viral (isolation & purification)</term>
<term>Antigens, Viral (analysis)</term>
<term>Enzyme-Linked Immunosorbent Assay (methods)</term>
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<term>SARS Virus (isolation & purification)</term>
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<term>Severe Acute Respiratory Syndrome (diagnosis)</term>
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<term>Adulte</term>
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<term>Anticorps monoclonaux (isolement et purification)</term>
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<term>Protéines nucléocapside (analyse)</term>
<term>Réaction de polymérisation en chaîne ()</term>
<term>Sensibilité et spécificité</term>
<term>Sujet âgé</term>
<term>Syndrome respiratoire aigu sévère (diagnostic)</term>
<term>Test ELISA ()</term>
<term>Virus du SRAS ()</term>
<term>Virus du SRAS (isolement et purification)</term>
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<term>Nucleocapsid Proteins</term>
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<term>Réaction de polymérisation en chaîne</term>
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<front><div type="abstract" xml:lang="en"><p>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.</p>
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<name sortKey="Fujimoto, Kotaro" sort="Fujimoto, Kotaro" uniqKey="Fujimoto K" first="Kotaro" last="Fujimoto">Kotaro Fujimoto</name>
<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>
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<name sortKey="Okamoto, Takashi" sort="Okamoto, Takashi" uniqKey="Okamoto T" first="Takashi" last="Okamoto">Takashi Okamoto</name>
<name sortKey="Takeda, Kazuhiko" sort="Takeda, Kazuhiko" uniqKey="Takeda K" first="Kazuhiko" last="Takeda">Kazuhiko Takeda</name>
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Serveur d'exploration SRAS

Sensitive and Specific Enzyme-Linked Immunosorbent Assay Using Chemiluminescence for Detection of Severe Acute Respiratory Syndrome Viral Infection▿

Sensitive and Specific Enzyme-Linked Immunosorbent Assay Using Chemiluminescence for Detection of Severe Acute Respiratory Syndrome Viral Infection▿

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Links to Exploration step

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

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