Evaluation of a safe and sensitive Spike protein-based immunofluorescence assay for the detection of antibody responses to SARS-CoV
Identifieur interne : 000D51 ( Ncbi/Curation ); précédent : 000D50; suivant : 000D52Evaluation of a safe and sensitive Spike protein-based immunofluorescence assay for the detection of antibody responses to SARS-CoV
Auteurs : Ivanus Manopo ; Liqun Lu ; Qigai He ; Li Lian Chee ; Shzu-Wei Chan ; Jimmy KwangSource :
- Journal of Immunological Methods [ 0022-1759 ] ; 2004.
Descripteurs français
- KwdFr :
- Animaux, Anticorps antiviraux (immunologie), Anticorps antiviraux (sang), Antigènes viraux (génétique), Antigènes viraux (immunologie), Baculoviridae (génétique), Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (immunologie), Humains, Microscopie de fluorescence, Protéines de l'enveloppe virale (immunologie), Protéines virales (génétique), Protéines virales (immunologie), Réactions croisées (immunologie), Sensibilité et spécificité, Spodoptera (cytologie), Spodoptera (génétique), Syndrome respiratoire aigu sévère (diagnostic), Technique d'immunofluorescence indirecte, Virus du SRAS (immunologie).
- MESH :
- cytologie : Spodoptera.
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Antigènes viraux, Baculoviridae, Protéines virales, Spodoptera.
- immunologie : Anticorps antiviraux, Antigènes viraux, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines virales, Réactions croisées, Virus du SRAS.
- sang : Anticorps antiviraux.
- Animaux, Glycoprotéine de spicule des coronavirus, Humains, Microscopie de fluorescence, Sensibilité et spécificité, Technique d'immunofluorescence indirecte.
English descriptors
- KwdEn :
- Animals, Antibodies, Viral (blood), Antibodies, Viral (immunology), Antigens, Viral (genetics), Antigens, Viral (immunology), Baculoviridae (genetics), Cross Reactions (immunology), Fluorescent Antibody Technique, Indirect, Humans, Membrane Glycoproteins (immunology), Microscopy, Fluorescence, SARS Virus (immunology), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), Spike Glycoprotein, Coronavirus, Spodoptera (cytology), Spodoptera (genetics), Viral Envelope Proteins (immunology), Viral Proteins (genetics), Viral Proteins (immunology).
- MESH :
- chemical , blood : Antibodies, Viral.
- chemical , genetics : Antigens, Viral, Viral Proteins.
- chemical , immunology : Antibodies, Viral, Antigens, Viral, Membrane Glycoproteins, Viral Envelope Proteins, Viral Proteins.
- cytology : Spodoptera.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : Baculoviridae, Spodoptera.
- immunology : Cross Reactions, SARS Virus.
- Animals, Fluorescent Antibody Technique, Indirect, Humans, Microscopy, Fluorescence, Sensitivity and Specificity, Spike Glycoprotein, Coronavirus.
Abstract
Previously, we have identified a truncated antigenic fragment named protein C [441 to 700 amino acids (a.a.)] as the immunodominant fragment of Spike (S) protein of severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV). We have now successfully expressed protein C using the baculovirus system in
Url:
DOI: 10.1016/j.jim.2004.10.012
PubMed: 15680149
PubMed Central: 7094334
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PMC:7094334Le document en format XML
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<author><name sortKey="Lu, Liqun" sort="Lu, Liqun" uniqKey="Lu L" first="Liqun" last="Lu">Liqun Lu</name>
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<author><name sortKey="He, Qigai" sort="He, Qigai" uniqKey="He Q" first="Qigai" last="He">Qigai He</name>
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<author><name sortKey="Chee, Li Lian" sort="Chee, Li Lian" uniqKey="Chee L" first="Li Lian" last="Chee">Li Lian Chee</name>
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<author><name sortKey="Chan, Shzu Wei" sort="Chan, Shzu Wei" uniqKey="Chan S" first="Shzu-Wei" last="Chan">Shzu-Wei Chan</name>
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<author><name sortKey="Kwang, Jimmy" sort="Kwang, Jimmy" uniqKey="Kwang J" first="Jimmy" last="Kwang">Jimmy Kwang</name>
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<series><title level="j">Journal of Immunological Methods</title>
<idno type="ISSN">0022-1759</idno>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term>
<term>Antibodies, Viral (blood)</term>
<term>Antibodies, Viral (immunology)</term>
<term>Antigens, Viral (genetics)</term>
<term>Antigens, Viral (immunology)</term>
<term>Baculoviridae (genetics)</term>
<term>Cross Reactions (immunology)</term>
<term>Fluorescent Antibody Technique, Indirect</term>
<term>Humans</term>
<term>Membrane Glycoproteins (immunology)</term>
<term>Microscopy, Fluorescence</term>
<term>SARS Virus (immunology)</term>
<term>Sensitivity and Specificity</term>
<term>Severe Acute Respiratory Syndrome (diagnosis)</term>
<term>Spike Glycoprotein, Coronavirus</term>
<term>Spodoptera (cytology)</term>
<term>Spodoptera (genetics)</term>
<term>Viral Envelope Proteins (immunology)</term>
<term>Viral Proteins (genetics)</term>
<term>Viral Proteins (immunology)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term>
<term>Anticorps antiviraux (immunologie)</term>
<term>Anticorps antiviraux (sang)</term>
<term>Antigènes viraux (génétique)</term>
<term>Antigènes viraux (immunologie)</term>
<term>Baculoviridae (génétique)</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Glycoprotéines membranaires (immunologie)</term>
<term>Humains</term>
<term>Microscopie de fluorescence</term>
<term>Protéines de l'enveloppe virale (immunologie)</term>
<term>Protéines virales (génétique)</term>
<term>Protéines virales (immunologie)</term>
<term>Réactions croisées (immunologie)</term>
<term>Sensibilité et spécificité</term>
<term>Spodoptera (cytologie)</term>
<term>Spodoptera (génétique)</term>
<term>Syndrome respiratoire aigu sévère (diagnostic)</term>
<term>Technique d'immunofluorescence indirecte</term>
<term>Virus du SRAS (immunologie)</term>
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</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Antigens, Viral</term>
<term>Viral Proteins</term>
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<keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Viral</term>
<term>Antigens, Viral</term>
<term>Membrane Glycoproteins</term>
<term>Viral Envelope Proteins</term>
<term>Viral Proteins</term>
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<keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term>
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<keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term>
</keywords>
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<term>Spodoptera</term>
</keywords>
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<term>Protéines virales</term>
<term>Spodoptera</term>
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<term>Antigènes viraux</term>
<term>Glycoprotéines membranaires</term>
<term>Protéines de l'enveloppe virale</term>
<term>Protéines virales</term>
<term>Réactions croisées</term>
<term>Virus du SRAS</term>
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<term>SARS Virus</term>
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<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Fluorescent Antibody Technique, Indirect</term>
<term>Humans</term>
<term>Microscopy, Fluorescence</term>
<term>Sensitivity and Specificity</term>
<term>Spike Glycoprotein, Coronavirus</term>
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<term>Glycoprotéine de spicule des coronavirus</term>
<term>Humains</term>
<term>Microscopie de fluorescence</term>
<term>Sensibilité et spécificité</term>
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<front><div type="abstract" xml:lang="en"><p>Previously, we have identified a truncated antigenic fragment named protein C [441 to 700 amino acids (a.a.)] as the immunodominant fragment of Spike (S) protein of severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV). We have now successfully expressed protein C using the baculovirus system in <italic>S. frugiperda</italic>
(Sf-9) cells. This recombinant baculovirus expressing protein C was first characterized using five SARS convalescent human sera and five normal human sera. The results showed that protein C is an authentic antigen against SARS-CoV antibody. Our Spike protein-based immunoflourescence assay (IFA) based on this recombinant baculovirus-Sf-9 system was further assessed with a panel of 163 clinical samples collected during the SARS epidemic in Singapore, which include samples from 21 clinically confirmed SARS, 42 non-SARS patient sera, and 100 normal sera. The results were compared to a commercial SARS IFA kit (EUROIMMUN, Germany) and a conventional IFA test performed in Singapore General Hospital. All of the 21 SARS-positive serum samples could be recognized by our IFA, giving a specificity and sensitivity of 100%, which was compatible with both whole virus-based IFA assays. No cross-reactivity with serum samples against infectious bronchitis virus (IBV) and transmissible gastroenteritis virus (TGEV) were detected in our assays. Thus, our Spike protein-based IFA could offer a safer procedure which can be performed in a BSL-2 laboratory as it could mimic the whole virus based-IFA without any loss of sensitivity and specificity. It is also more user-friendly and cost-effective than the whole virus-based IFA.</p>
</div>
</front>
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</TEI>
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