Comparison of two real-time quantitative assays for detection of severe acute respiratory syndrome coronavirus
Identifieur interne : 005B63 ( Main/Exploration ); précédent : 005B62; suivant : 005B64Comparison of two real-time quantitative assays for detection of severe acute respiratory syndrome coronavirus
Auteurs : Michael K. Hourfar [Allemagne] ; W. Kurt Roth [Allemagne] ; Erhard Seifried [Allemagne] ; Michael Schmidt [Allemagne]Source :
- Journal of clinical microbiology : (Print) [ 0095-1137 ] ; 2004.
Descripteurs français
- KwdFr :
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : ARN viral, Virus du SRAS.
- isolement et purification : Virus du SRAS.
- sang : ARN viral.
- virologie : Syndrome respiratoire aigu sévère.
- Pascal (Inist)
- Wicri :
- topic : Analyse quantitative.
English descriptors
- KwdEn :
- Coronavirus, Detection, Humans, Microbiology, Quantitative analysis, RNA, Viral (blood), RNA, Viral (genetics), Real time, Reverse Transcriptase Polymerase Chain Reaction (methods), Reverse Transcriptase Polymerase Chain Reaction (statistics & numerical data), SARS Virus (genetics), SARS Virus (isolation & purification), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (virology), Severe acute respiratory syndrome, Virology (methods), Virology (statistics & numerical data).
- MESH :
- chemical , blood : RNA, Viral.
- chemical , genetics : RNA, Viral.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- isolation & purification : SARS Virus.
- methods : Reverse Transcriptase Polymerase Chain Reaction, Virology.
- statistics & numerical data : Reverse Transcriptase Polymerase Chain Reaction, Virology.
- virology : Severe Acute Respiratory Syndrome.
- Humans, Sensitivity and Specificity.
Abstract
The new severe acute respiratory syndrome (SARS) coronavirus (CoV), described in February 2003, infected a total of 8,439 people. A total of 812 people died due to respiratory insufficiency. Close contact with symptomatic patients appeared to be the main route of transmission. However, potential transmission by blood transfusion could not be definitely excluded. Two real-time SARS-specific PCR assays were assessed for their sensitivities, agreement of test results, and intra-assay variabilities. Both assays rely on reverse transcription and amplification of extracted RNA. Dilutions of gamma-irradiated cell culture supernatants of SARS CoV-infected Vero E6 cells were prepared to determine the precisions, linear ranges, and accuracies of the assays. The linear range for the Artus RealArt HPA-Coronavirus assay (Artus assay) was 1 x 102 to 1 x 107 copies/ml, and that for the Roche LightCycler SARS CoV Quantification kit (Roche assay) was 1 x 104 to 2 x 108 copies/ml. The detection limit of the Roche assay was 3,982.1 copies/ml, whereas that of the Artus assay was 37.8 copies/ml. Detection limits were calculated with a standard preparation that was recommended for use by the World Health Organization. However, quantification of CoV in this preparation may be imprecise. In , both assays are suitable for quantitative measurement of SARS CoV at the high concentrations expected in sputum samples. The Artus assay is also suitable for detection of SARS CoV at the low concentrations found in serum samples.
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Affiliations:
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Hourfar</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Institute of Transfusion Medicine and Immunohematology, German Red Cross, Johann Wolfgang Goethe University</s1><s2>Frankfurt</s2><s3>DEU</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>Allemagne</country><placeName><region type="land" nuts="1">Hesse (Land)</region><region type="district" nuts="2">District de Darmstadt</region><settlement type="city">Francfort-sur-le-Main</settlement></placeName></affiliation></author><author><name sortKey="Roth, W Kurt" sort="Roth, W Kurt" uniqKey="Roth W" first="W. Kurt" last="Roth">W. Kurt Roth</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Institute of Transfusion Medicine and Immunohematology, German Red Cross, Johann Wolfgang Goethe University</s1><s2>Frankfurt</s2><s3>DEU</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>Allemagne</country><placeName><region type="land" nuts="1">Hesse (Land)</region><region type="district" nuts="2">District de Darmstadt</region><settlement type="city">Francfort-sur-le-Main</settlement></placeName></affiliation></author><author><name sortKey="Seifried, Erhard" sort="Seifried, Erhard" uniqKey="Seifried E" first="Erhard" last="Seifried">Erhard Seifried</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Institute of Transfusion Medicine and Immunohematology, German Red Cross, Johann Wolfgang Goethe University</s1><s2>Frankfurt</s2><s3>DEU</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>Allemagne</country><placeName><region type="land" nuts="1">Hesse (Land)</region><region type="district" nuts="2">District de Darmstadt</region><settlement type="city">Francfort-sur-le-Main</settlement></placeName></affiliation></author><author><name sortKey="Schmidt, Michael" sort="Schmidt, Michael" uniqKey="Schmidt M" first="Michael" last="Schmidt">Michael Schmidt</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Institute of Transfusion Medicine and Immunohematology, German Red Cross, Johann Wolfgang Goethe University</s1><s2>Frankfurt</s2><s3>DEU</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>Allemagne</country><placeName><region type="land" nuts="1">Hesse (Land)</region><region type="district" nuts="2">District de Darmstadt</region><settlement type="city">Francfort-sur-le-Main</settlement></placeName></affiliation></author></analytic><series><title level="j" type="main">Journal of clinical microbiology : (Print)</title><title level="j" type="abbreviated">J. clin. microbiol. : (Print)</title><idno type="ISSN">0095-1137</idno><imprint><date when="2004">2004</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of clinical microbiology : (Print)</title><title level="j" type="abbreviated">J. clin. microbiol. : (Print)</title><idno type="ISSN">0095-1137</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Coronavirus</term><term>Detection</term><term>Humans</term><term>Microbiology</term><term>Quantitative analysis</term><term>RNA, Viral (blood)</term><term>RNA, Viral (genetics)</term><term>Real time</term><term>Reverse Transcriptase Polymerase Chain Reaction (methods)</term><term>Reverse Transcriptase Polymerase Chain Reaction (statistics & numerical data)</term><term>SARS Virus (genetics)</term><term>SARS Virus (isolation & purification)</term><term>Sensitivity and Specificity</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term><term>Severe Acute Respiratory Syndrome (virology)</term><term>Severe acute respiratory syndrome</term><term>Virology (methods)</term><term>Virology (statistics & numerical data)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN viral (génétique)</term><term>ARN viral (sang)</term><term>Humains</term><term>RT-PCR ()</term><term>Sensibilité et spécificité</term><term>Syndrome respiratoire aigu sévère (diagnostic)</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Virologie ()</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (isolement et purification)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>RNA, Viral</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>RNA, Viral</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN viral</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>Virology</term></keywords><keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>ARN viral</term></keywords><keywords scheme="MESH" qualifier="statistics & numerical data" xml:lang="en"><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>Virology</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>Humans</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Coronavirus</term><term>Humains</term><term>RT-PCR</term><term>Sensibilité et spécificité</term><term>Temps réel</term><term>Analyse quantitative</term><term>Détection</term><term>Microbiologie</term><term>Syndrome respiratoire aigu sévère</term><term>Virologie</term></keywords><keywords scheme="Wicri" type="topic" xml:lang="fr"><term>Analyse quantitative</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The new severe acute respiratory syndrome (SARS) coronavirus (CoV), described in February 2003, infected a total of 8,439 people. A total of 812 people died due to respiratory insufficiency. Close contact with symptomatic patients appeared to be the main route of transmission. However, potential transmission by blood transfusion could not be definitely excluded. Two real-time SARS-specific PCR assays were assessed for their sensitivities, agreement of test results, and intra-assay variabilities. Both assays rely on reverse transcription and amplification of extracted RNA. Dilutions of gamma-irradiated cell culture supernatants of SARS CoV-infected Vero E6 cells were prepared to determine the precisions, linear ranges, and accuracies of the assays. The linear range for the Artus RealArt HPA-Coronavirus assay (Artus assay) was 1 x 10<sup>2</sup> to 1 x 10<sup>7</sup> copies/ml, and that for the Roche LightCycler SARS CoV Quantification kit (Roche assay) was 1 x 104 to 2 x 10<sup>8</sup> copies/ml. The detection limit of the Roche assay was 3,982.1 copies/ml, whereas that of the Artus assay was 37.8 copies/ml. Detection limits were calculated with a standard preparation that was recommended for use by the World Health Organization. However, quantification of CoV in this preparation may be imprecise. In , both assays are suitable for quantitative measurement of SARS CoV at the high concentrations expected in sputum samples. The Artus assay is also suitable for detection of SARS CoV at the low concentrations found in serum samples.</div></front></TEI><affiliations><list><country><li>Allemagne</li></country><region><li>District de Darmstadt</li><li>Hesse (Land)</li></region><settlement><li>Francfort-sur-le-Main</li></settlement></list><tree><country name="Allemagne"><region name="Hesse (Land)"><name sortKey="Hourfar, Michael K" sort="Hourfar, Michael K" uniqKey="Hourfar M" first="Michael K." last="Hourfar">Michael K. Hourfar</name></region><name sortKey="Roth, W Kurt" sort="Roth, W Kurt" uniqKey="Roth W" first="W. Kurt" last="Roth">W. Kurt Roth</name><name sortKey="Schmidt, Michael" sort="Schmidt, Michael" uniqKey="Schmidt M" first="Michael" last="Schmidt">Michael Schmidt</name><name sortKey="Seifried, Erhard" sort="Seifried, Erhard" uniqKey="Seifried E" first="Erhard" last="Seifried">Erhard Seifried</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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