Evaluation of a safe and sensitive Spike protein-based immunofluorescence assay for the detection of antibody responses to SARS-CoV
Identifieur interne : 004E30 ( Main/Exploration ); précédent : 004E29; suivant : 004E31Evaluation of a safe and sensitive Spike protein-based immunofluorescence assay for the detection of antibody responses to SARS-CoV
Auteurs : Ivanus Manopo [Singapour] ; LIQUN LU [Singapour] ; QIGAI HE [Singapour] ; LI LIAN CHEE [Singapour] ; Shzu-Wei Chan [Singapour] ; Jimmy Kwang [Singapour]Source :
- Journal of immunological methods [ 0022-1759 ] ; 2005.
Descripteurs français
- Pascal (Inist)
English descriptors
- KwdEn :
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 S. frugiperda (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.
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream PascalFrancis, to step Corpus: 000725
- to stream PascalFrancis, to step Curation: 000265
- to stream PascalFrancis, to step Checkpoint: 000655
- to stream Main, to step Merge: 005175
- to stream Main, to step Curation: 004E30
Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en" level="a">Evaluation of a safe and sensitive Spike protein-based immunofluorescence assay for the detection of antibody responses to SARS-CoV</title>
<author><name sortKey="Manopo, Ivanus" sort="Manopo, Ivanus" uniqKey="Manopo I" first="Ivanus" last="Manopo">Ivanus Manopo</name>
<affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Sciences Laboratory, 1 Research link, National University of singapore</s1>
<s2>117604 Singapore</s2>
<s3>SGP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
</inist:fA14>
<country>Singapour</country>
<orgName type="university">Université nationale de Singapour</orgName>
</affiliation>
</author>
<author><name sortKey="Liqun Lu" sort="Liqun Lu" uniqKey="Liqun Lu" last="Liqun Lu">LIQUN LU</name>
<affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Sciences Laboratory, 1 Research link, National University of singapore</s1>
<s2>117604 Singapore</s2>
<s3>SGP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
</inist:fA14>
<country>Singapour</country>
<orgName type="university">Université nationale de Singapour</orgName>
</affiliation>
</author>
<author><name sortKey="Qigai He" sort="Qigai He" uniqKey="Qigai He" last="Qigai He">QIGAI HE</name>
<affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Sciences Laboratory, 1 Research link, National University of singapore</s1>
<s2>117604 Singapore</s2>
<s3>SGP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
</inist:fA14>
<country>Singapour</country>
<orgName type="university">Université nationale de Singapour</orgName>
</affiliation>
</author>
<author><name sortKey="Li Lian Chee" sort="Li Lian Chee" uniqKey="Li Lian Chee" last="Li Lian Chee">LI LIAN CHEE</name>
<affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Sciences Laboratory, 1 Research link, National University of singapore</s1>
<s2>117604 Singapore</s2>
<s3>SGP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
</inist:fA14>
<country>Singapour</country>
<orgName type="university">Université nationale de Singapour</orgName>
</affiliation>
</author>
<author><name sortKey="Chan, Shzu Wei" sort="Chan, Shzu Wei" uniqKey="Chan S" first="Shzu-Wei" last="Chan">Shzu-Wei Chan</name>
<affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Sciences Laboratory, 1 Research link, National University of singapore</s1>
<s2>117604 Singapore</s2>
<s3>SGP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
</inist:fA14>
<country>Singapour</country>
<orgName type="university">Université nationale de Singapour</orgName>
</affiliation>
</author>
<author><name sortKey="Kwang, Jimmy" sort="Kwang, Jimmy" uniqKey="Kwang J" first="Jimmy" last="Kwang">Jimmy Kwang</name>
<affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Sciences Laboratory, 1 Research link, National University of singapore</s1>
<s2>117604 Singapore</s2>
<s3>SGP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
</inist:fA14>
<country>Singapour</country>
<orgName type="university">Université nationale de Singapour</orgName>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">INIST</idno>
<idno type="inist">05-0121836</idno>
<date when="2005">2005</date>
<idno type="stanalyst">PASCAL 05-0121836 INIST</idno>
<idno type="RBID">Pascal:05-0121836</idno>
<idno type="wicri:Area/PascalFrancis/Corpus">000725</idno>
<idno type="wicri:Area/PascalFrancis/Curation">000265</idno>
<idno type="wicri:Area/PascalFrancis/Checkpoint">000655</idno>
<idno type="wicri:explorRef" wicri:stream="PascalFrancis" wicri:step="Checkpoint">000655</idno>
<idno type="wicri:doubleKey">0022-1759:2005:Manopo I:evaluation:of:a</idno>
<idno type="wicri:Area/Main/Merge">005175</idno>
<idno type="wicri:Area/Main/Curation">004E30</idno>
<idno type="wicri:Area/Main/Exploration">004E30</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Evaluation of a safe and sensitive Spike protein-based immunofluorescence assay for the detection of antibody responses to SARS-CoV</title>
<author><name sortKey="Manopo, Ivanus" sort="Manopo, Ivanus" uniqKey="Manopo I" first="Ivanus" last="Manopo">Ivanus Manopo</name>
<affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Sciences Laboratory, 1 Research link, National University of singapore</s1>
<s2>117604 Singapore</s2>
<s3>SGP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
</inist:fA14>
<country>Singapour</country>
<orgName type="university">Université nationale de Singapour</orgName>
</affiliation>
</author>
<author><name sortKey="Liqun Lu" sort="Liqun Lu" uniqKey="Liqun Lu" last="Liqun Lu">LIQUN LU</name>
<affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Sciences Laboratory, 1 Research link, National University of singapore</s1>
<s2>117604 Singapore</s2>
<s3>SGP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
</inist:fA14>
<country>Singapour</country>
<orgName type="university">Université nationale de Singapour</orgName>
</affiliation>
</author>
<author><name sortKey="Qigai He" sort="Qigai He" uniqKey="Qigai He" last="Qigai He">QIGAI HE</name>
<affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Sciences Laboratory, 1 Research link, National University of singapore</s1>
<s2>117604 Singapore</s2>
<s3>SGP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
</inist:fA14>
<country>Singapour</country>
<orgName type="university">Université nationale de Singapour</orgName>
</affiliation>
</author>
<author><name sortKey="Li Lian Chee" sort="Li Lian Chee" uniqKey="Li Lian Chee" last="Li Lian Chee">LI LIAN CHEE</name>
<affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Sciences Laboratory, 1 Research link, National University of singapore</s1>
<s2>117604 Singapore</s2>
<s3>SGP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
</inist:fA14>
<country>Singapour</country>
<orgName type="university">Université nationale de Singapour</orgName>
</affiliation>
</author>
<author><name sortKey="Chan, Shzu Wei" sort="Chan, Shzu Wei" uniqKey="Chan S" first="Shzu-Wei" last="Chan">Shzu-Wei Chan</name>
<affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Sciences Laboratory, 1 Research link, National University of singapore</s1>
<s2>117604 Singapore</s2>
<s3>SGP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
</inist:fA14>
<country>Singapour</country>
<orgName type="university">Université nationale de Singapour</orgName>
</affiliation>
</author>
<author><name sortKey="Kwang, Jimmy" sort="Kwang, Jimmy" uniqKey="Kwang J" first="Jimmy" last="Kwang">Jimmy Kwang</name>
<affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Sciences Laboratory, 1 Research link, National University of singapore</s1>
<s2>117604 Singapore</s2>
<s3>SGP</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
</inist:fA14>
<country>Singapour</country>
<orgName type="university">Université nationale de Singapour</orgName>
</affiliation>
</author>
</analytic>
<series><title level="j" type="main">Journal of immunological methods</title>
<title level="j" type="abbreviated">J. immunol. methods</title>
<idno type="ISSN">0022-1759</idno>
<imprint><date when="2005">2005</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
<seriesStmt><title level="j" type="main">Journal of immunological methods</title>
<title level="j" type="abbreviated">J. immunol. methods</title>
<idno type="ISSN">0022-1759</idno>
</seriesStmt>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibody</term>
<term>Detection</term>
<term>Humoral immunity</term>
<term>Immunofluorescence</term>
<term>Immunological method</term>
<term>Protein</term>
<term>Severe acute respiratory syndrome</term>
</keywords>
<keywords scheme="Pascal" xml:lang="fr"><term>Protéine</term>
<term>Immunofluorescence</term>
<term>Détection</term>
<term>Immunité humorale</term>
<term>Anticorps</term>
<term>Méthode immunologique</term>
<term>Syndrome respiratoire aigu sévère</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">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 S. frugiperda (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.</div>
</front>
</TEI>
<affiliations><list><country><li>Singapour</li>
</country>
<orgName><li>Université nationale de Singapour</li>
</orgName>
</list>
<tree><country name="Singapour"><noRegion><name sortKey="Manopo, Ivanus" sort="Manopo, Ivanus" uniqKey="Manopo I" first="Ivanus" last="Manopo">Ivanus Manopo</name>
</noRegion>
<name sortKey="Chan, Shzu Wei" sort="Chan, Shzu Wei" uniqKey="Chan S" first="Shzu-Wei" last="Chan">Shzu-Wei Chan</name>
<name sortKey="Kwang, Jimmy" sort="Kwang, Jimmy" uniqKey="Kwang J" first="Jimmy" last="Kwang">Jimmy Kwang</name>
<name sortKey="Li Lian Chee" sort="Li Lian Chee" uniqKey="Li Lian Chee" last="Li Lian Chee">LI LIAN CHEE</name>
<name sortKey="Liqun Lu" sort="Liqun Lu" uniqKey="Liqun Lu" last="Liqun Lu">LIQUN LU</name>
<name sortKey="Qigai He" sort="Qigai He" uniqKey="Qigai He" last="Qigai He">QIGAI HE</name>
</country>
</tree>
</affiliations>
</record>
Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/SrasV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 004E30 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 004E30 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien |wiki= Sante |area= SrasV1 |flux= Main |étape= Exploration |type= RBID |clé= Pascal:05-0121836 |texte= Evaluation of a safe and sensitive Spike protein-based immunofluorescence assay for the detection of antibody responses to SARS-CoV }}
![]() | This area was generated with Dilib version V0.6.33. | ![]() |