Novel rapid immunochromatographic test based on an enzyme immunoassay for detecting nucleocapsid antigen in SARS-associated coronavirus.
Identifieur interne : 004738 ( Main/Exploration ); précédent : 004737; suivant : 004739Novel rapid immunochromatographic test based on an enzyme immunoassay for detecting nucleocapsid antigen in SARS-associated coronavirus.
Auteurs : Hiroyuki Kogaki [Japon] ; Yoshiaki Uchida ; Nobuyuki Fujii ; Yoshihiro Kurano ; Kazushige Miyake ; Yasuji Kido ; Hiroaki Kariwa ; Ikuo Takashima ; Hiko Tamashiro ; Ai-Ee Ling ; Masahisa OkadaSource :
- Journal of clinical laboratory analysis [ 0887-8013 ] ; 2005.
Descripteurs français
- KwdFr :
- ADN recombiné, Animaux, Anticorps monoclonaux (immunologie), Cellules Vero, Chromatographie, Coronavirus (immunologie), Escherichia coli, Facteurs temps, Nucléocapside (analyse), Nucléocapside (immunologie), Reproductibilité des résultats, Sensibilité et spécificité, Souris, Syndrome respiratoire aigu sévère (virologie), Techniques immunoenzymatiques (), Techniques immunoenzymatiques (instrumentation).
- MESH :
- analyse : Nucléocapside.
- immunologie : Anticorps monoclonaux, Coronavirus, Nucléocapside.
- virologie : Syndrome respiratoire aigu sévère.
- instrumentation : ADN recombiné, Animaux, Cellules Vero, Chromatographie, Escherichia coli, Facteurs temps, Reproductibilité des résultats, Sensibilité et spécificité, Souris, Techniques immunoenzymatiques.
English descriptors
- KwdEn :
- Animals, Antibodies, Monoclonal (immunology), Chlorocebus aethiops, Chromatography, Coronavirus (immunology), DNA, Recombinant, Escherichia coli, Immunoenzyme Techniques (instrumentation), Immunoenzyme Techniques (methods), Mice, Nucleocapsid (analysis), Nucleocapsid (immunology), Reproducibility of Results, Sensitivity and Specificity, Severe Acute Respiratory Syndrome (virology), Time Factors, Vero Cells.
- MESH :
- chemical , immunology : Antibodies, Monoclonal.
- analysis : Nucleocapsid.
- immunology : Coronavirus, Nucleocapsid.
- instrumentation : Immunoenzyme Techniques.
- methods : Immunoenzyme Techniques.
- virology : Severe Acute Respiratory Syndrome.
- Animals, Chlorocebus aethiops, Chromatography, DNA, Recombinant, Escherichia coli, Mice, Reproducibility of Results, Sensitivity and Specificity, Time Factors, Vero Cells.
Abstract
A novel severe acute respiratory syndrome-associated coronavirus (SARS-CoV) has been discovered. The detection of both antigens and antibodies in SARS-CoV from human specimens with suspected SARS plays an important role in preventing infection. We developed a novel rapid immunochromatographic test (RICT) based on the sandwich format enzyme immunoassay (EIA) with an all-in-one device for detecting the native nucleocapsid antigen (N-Ag) of SARS-CoV using monoclonal antibodies (MoAbs), which we produced by immunizing recombinant N-Ag to mice. RICT is a qualitative assay for respiratory aspirates and serum specimens. With this assay, a positive result can be judged subjectively by the appearance of a blue line on the device 15 min after the sample is applied. RICT with several pairs of MoAbs showed a high sensitivity for the detection of recombinant N-Ag as well as viral N-Ag of SARS-CoV. rSN122 and rSN21-2 were the best MoAbs for immobilized antibody and enzyme labeling, respectively. With regard to analytical sensitivity, RICT detected N-Ag at 31 pg/mL for recombinant N-Ag, and at 1.99 x 10(2) TCID(50)/mL for SARS-CoV. The specificity of RICT was 100% when 150 human sera and 50 nasopharyngeal aspirates (NSPs) were used. RICT based on an EIA using the rSN122/rSN21-2 pair is a sensitive, specific, and reliable rapid assay for detecting N-Ag in SARS-CoV treated with either heat or Triton X-100.
DOI: 10.1002/jcla.20070
PubMed: 16025480
Affiliations:
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sortKey="Uchida, Yoshiaki" sort="Uchida, Yoshiaki" uniqKey="Uchida Y" first="Yoshiaki" last="Uchida">Yoshiaki Uchida</name></author><author><name sortKey="Fujii, Nobuyuki" sort="Fujii, Nobuyuki" uniqKey="Fujii N" first="Nobuyuki" last="Fujii">Nobuyuki Fujii</name></author><author><name sortKey="Kurano, Yoshihiro" sort="Kurano, Yoshihiro" uniqKey="Kurano Y" first="Yoshihiro" last="Kurano">Yoshihiro Kurano</name></author><author><name sortKey="Miyake, Kazushige" sort="Miyake, Kazushige" uniqKey="Miyake K" first="Kazushige" last="Miyake">Kazushige Miyake</name></author><author><name sortKey="Kido, Yasuji" sort="Kido, Yasuji" uniqKey="Kido Y" first="Yasuji" last="Kido">Yasuji Kido</name></author><author><name sortKey="Kariwa, Hiroaki" sort="Kariwa, Hiroaki" uniqKey="Kariwa H" first="Hiroaki" last="Kariwa">Hiroaki Kariwa</name></author><author><name sortKey="Takashima, Ikuo" sort="Takashima, Ikuo" uniqKey="Takashima I" first="Ikuo" last="Takashima">Ikuo Takashima</name></author><author><name sortKey="Tamashiro, Hiko" sort="Tamashiro, Hiko" uniqKey="Tamashiro H" first="Hiko" last="Tamashiro">Hiko Tamashiro</name></author><author><name sortKey="Ling, Ai Ee" sort="Ling, Ai Ee" uniqKey="Ling A" first="Ai-Ee" last="Ling">Ai-Ee Ling</name></author><author><name sortKey="Okada, Masahisa" sort="Okada, Masahisa" uniqKey="Okada M" first="Masahisa" last="Okada">Masahisa Okada</name></author></analytic><series><title level="j">Journal of clinical laboratory analysis</title><idno type="ISSN">0887-8013</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antibodies, Monoclonal (immunology)</term><term>Chlorocebus aethiops</term><term>Chromatography</term><term>Coronavirus (immunology)</term><term>DNA, Recombinant</term><term>Escherichia coli</term><term>Immunoenzyme Techniques (instrumentation)</term><term>Immunoenzyme Techniques (methods)</term><term>Mice</term><term>Nucleocapsid (analysis)</term><term>Nucleocapsid (immunology)</term><term>Reproducibility of Results</term><term>Sensitivity and Specificity</term><term>Severe Acute Respiratory Syndrome (virology)</term><term>Time Factors</term><term>Vero Cells</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN recombiné</term><term>Animaux</term><term>Anticorps monoclonaux (immunologie)</term><term>Cellules Vero</term><term>Chromatographie</term><term>Coronavirus (immunologie)</term><term>Escherichia coli</term><term>Facteurs temps</term><term>Nucléocapside (analyse)</term><term>Nucléocapside (immunologie)</term><term>Reproductibilité des résultats</term><term>Sensibilité et spécificité</term><term>Souris</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Techniques immunoenzymatiques ()</term><term>Techniques immunoenzymatiques (instrumentation)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Monoclonal</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Nucléocapside</term></keywords><keywords scheme="MESH" qualifier="analysis" xml:lang="en"><term>Nucleocapsid</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps monoclonaux</term><term>Coronavirus</term><term>Nucléocapside</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Coronavirus</term><term>Nucleocapsid</term></keywords><keywords scheme="MESH" qualifier="instrumentation" xml:lang="en"><term>Immunoenzyme Techniques</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Immunoenzyme Techniques</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>Chromatography</term><term>DNA, Recombinant</term><term>Escherichia coli</term><term>Mice</term><term>Reproducibility of Results</term><term>Sensitivity and Specificity</term><term>Time Factors</term><term>Vero Cells</term></keywords><keywords scheme="MESH" qualifier="instrumentation" xml:lang="fr"><term>ADN recombiné</term><term>Animaux</term><term>Cellules Vero</term><term>Chromatographie</term><term>Escherichia coli</term><term>Facteurs temps</term><term>Reproductibilité des résultats</term><term>Sensibilité et spécificité</term><term>Souris</term><term>Techniques immunoenzymatiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A novel severe acute respiratory syndrome-associated coronavirus (SARS-CoV) has been discovered. The detection of both antigens and antibodies in SARS-CoV from human specimens with suspected SARS plays an important role in preventing infection. We developed a novel rapid immunochromatographic test (RICT) based on the sandwich format enzyme immunoassay (EIA) with an all-in-one device for detecting the native nucleocapsid antigen (N-Ag) of SARS-CoV using monoclonal antibodies (MoAbs), which we produced by immunizing recombinant N-Ag to mice. RICT is a qualitative assay for respiratory aspirates and serum specimens. With this assay, a positive result can be judged subjectively by the appearance of a blue line on the device 15 min after the sample is applied. RICT with several pairs of MoAbs showed a high sensitivity for the detection of recombinant N-Ag as well as viral N-Ag of SARS-CoV. rSN122 and rSN21-2 were the best MoAbs for immobilized antibody and enzyme labeling, respectively. With regard to analytical sensitivity, RICT detected N-Ag at 31 pg/mL for recombinant N-Ag, and at 1.99 x 10(2) TCID(50)/mL for SARS-CoV. The specificity of RICT was 100% when 150 human sera and 50 nasopharyngeal aspirates (NSPs) were used. RICT based on an EIA using the rSN122/rSN21-2 pair is a sensitive, specific, and reliable rapid assay for detecting N-Ag in SARS-CoV treated with either heat or Triton X-100.</div></front></TEI><affiliations><list><country><li>Japon</li></country><region><li>Région de Kantō</li></region><settlement><li>Tokyo</li></settlement></list><tree><noCountry><name sortKey="Fujii, Nobuyuki" sort="Fujii, Nobuyuki" uniqKey="Fujii N" first="Nobuyuki" last="Fujii">Nobuyuki Fujii</name><name sortKey="Kariwa, Hiroaki" sort="Kariwa, Hiroaki" uniqKey="Kariwa H" first="Hiroaki" last="Kariwa">Hiroaki Kariwa</name><name sortKey="Kido, Yasuji" sort="Kido, Yasuji" uniqKey="Kido Y" first="Yasuji" last="Kido">Yasuji Kido</name><name sortKey="Kurano, Yoshihiro" sort="Kurano, Yoshihiro" uniqKey="Kurano Y" first="Yoshihiro" last="Kurano">Yoshihiro Kurano</name><name sortKey="Ling, Ai Ee" sort="Ling, Ai Ee" uniqKey="Ling A" first="Ai-Ee" last="Ling">Ai-Ee Ling</name><name sortKey="Miyake, Kazushige" sort="Miyake, Kazushige" uniqKey="Miyake K" first="Kazushige" last="Miyake">Kazushige Miyake</name><name sortKey="Okada, Masahisa" sort="Okada, Masahisa" uniqKey="Okada M" first="Masahisa" last="Okada">Masahisa Okada</name><name sortKey="Takashima, Ikuo" sort="Takashima, Ikuo" uniqKey="Takashima I" first="Ikuo" last="Takashima">Ikuo Takashima</name><name sortKey="Tamashiro, Hiko" sort="Tamashiro, Hiko" uniqKey="Tamashiro H" first="Hiko" last="Tamashiro">Hiko Tamashiro</name><name sortKey="Uchida, Yoshiaki" sort="Uchida, Yoshiaki" uniqKey="Uchida Y" first="Yoshiaki" last="Uchida">Yoshiaki Uchida</name></noCountry><country name="Japon"><region name="Région de Kantō"><name sortKey="Kogaki, Hiroyuki" sort="Kogaki, Hiroyuki" uniqKey="Kogaki H" first="Hiroyuki" last="Kogaki">Hiroyuki Kogaki</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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