Evaluation of a safe and sensitive Spike protein-based immunofluorescence assay for the detection of antibody responses to SARS-CoV
Identifieur interne : 000725 ( PascalFrancis/Corpus ); précédent : 000724; suivant : 000726Evaluation 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 ] ; 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.
Notice en format standard (ISO 2709)
Pour connaître la documentation sur le format Inist Standard.
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Format Inist (serveur)
NO : | PASCAL 05-0121836 INIST |
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ET : | Evaluation of a safe and sensitive Spike protein-based immunofluorescence assay for the detection of antibody responses to SARS-CoV |
AU : | MANOPO (Ivanus); LIQUN LU; QIGAI HE; LI LIAN CHEE; CHAN (Shzu-Wei); KWANG (Jimmy) |
AF : | Animal Health Biotechnology, Temasek Life Sciences Laboratory, 1 Research link, National University of singapore/117604 Singapore/Singapour (1 aut., 2 aut., 3 aut., 4 aut., 5 aut., 6 aut.) |
DT : | Publication en série; Papier de recherche; Niveau analytique |
SO : | Journal of immunological methods; ISSN 0022-1759; Coden JIMMBG; Pays-Bas; Da. 2005; Vol. 296; No. 1-2; Pp. 37-44; Bibl. 18 ref. |
LA : | Anglais |
EA : | 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. |
CC : | 002A06B06 |
FD : | Protéine; Immunofluorescence; Détection; Immunité humorale; Anticorps; Méthode immunologique; Syndrome respiratoire aigu sévère |
FG : | Virose; Infection |
ED : | Protein; Immunofluorescence; Detection; Humoral immunity; Antibody; Immunological method; Severe acute respiratory syndrome |
EG : | Viral disease; Infection |
SD : | Proteína; Inmunofluorescencia; Detección; Inmunidad humoral; Anticuerpo; Método inmunológico; Síndrome respiratorio agudo severo |
LO : | INIST-15654.354000126735630050 |
ID : | 05-0121836 |
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Pascal:05-0121836Le document en format XML
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<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>
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<server><NO>PASCAL 05-0121836 INIST</NO>
<ET>Evaluation of a safe and sensitive Spike protein-based immunofluorescence assay for the detection of antibody responses to SARS-CoV</ET>
<AU>MANOPO (Ivanus); LIQUN LU; QIGAI HE; LI LIAN CHEE; CHAN (Shzu-Wei); KWANG (Jimmy)</AU>
<AF>Animal Health Biotechnology, Temasek Life Sciences Laboratory, 1 Research link, National University of singapore/117604 Singapore/Singapour (1 aut., 2 aut., 3 aut., 4 aut., 5 aut., 6 aut.)</AF>
<DT>Publication en série; Papier de recherche; Niveau analytique</DT>
<SO>Journal of immunological methods; ISSN 0022-1759; Coden JIMMBG; Pays-Bas; Da. 2005; Vol. 296; No. 1-2; Pp. 37-44; Bibl. 18 ref.</SO>
<LA>Anglais</LA>
<EA>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.</EA>
<CC>002A06B06</CC>
<FD>Protéine; Immunofluorescence; Détection; Immunité humorale; Anticorps; Méthode immunologique; Syndrome respiratoire aigu sévère</FD>
<FG>Virose; Infection</FG>
<ED>Protein; Immunofluorescence; Detection; Humoral immunity; Antibody; Immunological method; Severe acute respiratory syndrome</ED>
<EG>Viral disease; Infection</EG>
<SD>Proteína; Inmunofluorescencia; Detección; Inmunidad humoral; Anticuerpo; Método inmunológico; Síndrome respiratorio agudo severo</SD>
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<ID>05-0121836</ID>
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