A real-time PCR for SARS-coronavirus incorporating target gene pre-amplification.
Identifieur interne : 006093 ( Main/Exploration ); précédent : 006092; suivant : 006094A real-time PCR for SARS-coronavirus incorporating target gene pre-amplification.
Auteurs : Lok Ting Lau [République populaire de Chine] ; Yin-Wan Wendy Fung ; Freda Pui-Fan Wong ; Selma Sau-Wah Lin ; Chen Ran Wang ; Hui Li Li ; Natalie Dillon ; Richard A. Collins ; John Siu-Lun Tam ; Paul K S. Chan ; Chen G. Wang ; Albert Cheung-Hoi YuSource :
- Biochemical and biophysical research communications [ 0006-291X ] ; 2003.
Descripteurs français
- KwdFr :
- Dépistage génétique (), Génome viral, Humains, Reproductibilité des résultats, Réaction de polymérisation en chaîne (), Sensibilité et spécificité, Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (virologie), Systèmes en direct, Techniques d'amplification d'acides nucléiques (), Virus du SRAS (génétique), Virus du SRAS (isolement et purification).
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Virus du SRAS.
- isolement et purification : Virus du SRAS.
- virologie : Syndrome respiratoire aigu sévère.
- Dépistage génétique, Génome viral, Humains, Reproductibilité des résultats, Réaction de polymérisation en chaîne, Sensibilité et spécificité, Systèmes en direct, Techniques d'amplification d'acides nucléiques.
English descriptors
- KwdEn :
- Genetic Testing (methods), Genome, Viral, Humans, Nucleic Acid Amplification Techniques (methods), Online Systems, Polymerase Chain Reaction (methods), Reproducibility of Results, SARS Virus (genetics), SARS Virus (isolation & purification), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (virology).
- MESH :
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- isolation & purification : SARS Virus.
- methods : Genetic Testing, Nucleic Acid Amplification Techniques, Polymerase Chain Reaction.
- virology : Severe Acute Respiratory Syndrome.
- Genome, Viral, Humans, Online Systems, Reproducibility of Results, Sensitivity and Specificity.
Abstract
An enhanced polymerase chain reaction (PCR) assay to detect the coronavirus associated with severe acute respiratory syndrome (SARS-CoV) was developed in which a target gene pre-amplification step preceded TaqMan real-time fluorescent PCR. Clinical samples were collected from 120 patients diagnosed as suspected or probable SARS cases and analyzed by conventional PCR followed by agarose gel electrophoresis, conventional TaqMan real-time PCR, and our enhanced TaqMan real-time PCR assays. An amplicon of the size expected from SARS-CoV was obtained from 28/120 samples using the enhanced real-time PCR method. Conventional PCR and real-time PCR alone identified fewer SARS-CoV positive cases. Results were confirmed by viral culture in 3/28 cases. The limit of detection of the enhanced real-time PCR method was 10(2)-fold higher than the standard real-time PCR assay and 10(7)-fold higher than conventional PCR methods. The increased sensitivity of the assay may help control the spread of the disease during future SARS outbreaks.
DOI: 10.1016/j.bbrc.2003.11.064
PubMed: 14652014
Affiliations:
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Le document en format XML
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Collins</name></author><author><name sortKey="Tam, John Siu Lun" sort="Tam, John Siu Lun" uniqKey="Tam J" first="John Siu-Lun" last="Tam">John Siu-Lun Tam</name></author><author><name sortKey="Chan, Paul K S" sort="Chan, Paul K S" uniqKey="Chan P" first="Paul K S" last="Chan">Paul K S. Chan</name></author><author><name sortKey="Wang, Chen G" sort="Wang, Chen G" uniqKey="Wang C" first="Chen G" last="Wang">Chen G. Wang</name></author><author><name sortKey="Yu, Albert Cheung Hoi" sort="Yu, Albert Cheung Hoi" uniqKey="Yu A" first="Albert Cheung-Hoi" last="Yu">Albert Cheung-Hoi Yu</name></author></analytic><series><title level="j">Biochemical and biophysical research communications</title><idno type="ISSN">0006-291X</idno><imprint><date when="2003" type="published">2003</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Genetic Testing (methods)</term><term>Genome, Viral</term><term>Humans</term><term>Nucleic Acid Amplification Techniques (methods)</term><term>Online Systems</term><term>Polymerase Chain Reaction (methods)</term><term>Reproducibility of Results</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></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Dépistage génétique ()</term><term>Génome viral</term><term>Humains</term><term>Reproductibilité des résultats</term><term>Réaction de polymérisation en chaîne ()</term><term>Sensibilité et spécificité</term><term>Syndrome respiratoire aigu sévère (diagnostic)</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Systèmes en direct</term><term>Techniques d'amplification d'acides nucléiques ()</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (isolement et purification)</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>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>Genetic Testing</term><term>Nucleic Acid Amplification Techniques</term><term>Polymerase Chain Reaction</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>Genome, Viral</term><term>Humans</term><term>Online Systems</term><term>Reproducibility of Results</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Dépistage génétique</term><term>Génome viral</term><term>Humains</term><term>Reproductibilité des résultats</term><term>Réaction de polymérisation en chaîne</term><term>Sensibilité et spécificité</term><term>Systèmes en direct</term><term>Techniques d'amplification d'acides nucléiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">An enhanced polymerase chain reaction (PCR) assay to detect the coronavirus associated with severe acute respiratory syndrome (SARS-CoV) was developed in which a target gene pre-amplification step preceded TaqMan real-time fluorescent PCR. Clinical samples were collected from 120 patients diagnosed as suspected or probable SARS cases and analyzed by conventional PCR followed by agarose gel electrophoresis, conventional TaqMan real-time PCR, and our enhanced TaqMan real-time PCR assays. An amplicon of the size expected from SARS-CoV was obtained from 28/120 samples using the enhanced real-time PCR method. Conventional PCR and real-time PCR alone identified fewer SARS-CoV positive cases. Results were confirmed by viral culture in 3/28 cases. The limit of detection of the enhanced real-time PCR method was 10(2)-fold higher than the standard real-time PCR assay and 10(7)-fold higher than conventional PCR methods. The increased sensitivity of the assay may help control the spread of the disease during future SARS outbreaks.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><settlement><li>Pékin</li></settlement></list><tree><noCountry><name sortKey="Chan, Paul K S" sort="Chan, Paul K S" uniqKey="Chan P" first="Paul K S" last="Chan">Paul K S. Chan</name><name sortKey="Collins, Richard A" sort="Collins, Richard A" uniqKey="Collins R" first="Richard A" last="Collins">Richard A. Collins</name><name sortKey="Dillon, Natalie" sort="Dillon, Natalie" uniqKey="Dillon N" first="Natalie" last="Dillon">Natalie Dillon</name><name sortKey="Fung, Yin Wan Wendy" sort="Fung, Yin Wan Wendy" uniqKey="Fung Y" first="Yin-Wan Wendy" last="Fung">Yin-Wan Wendy Fung</name><name sortKey="Li, Hui Li" sort="Li, Hui Li" uniqKey="Li H" first="Hui Li" last="Li">Hui Li Li</name><name sortKey="Lin, Selma Sau Wah" sort="Lin, Selma Sau Wah" uniqKey="Lin S" first="Selma Sau-Wah" last="Lin">Selma Sau-Wah Lin</name><name sortKey="Tam, John Siu Lun" sort="Tam, John Siu Lun" uniqKey="Tam J" first="John Siu-Lun" last="Tam">John Siu-Lun Tam</name><name sortKey="Wang, Chen G" sort="Wang, Chen G" uniqKey="Wang C" first="Chen G" last="Wang">Chen G. Wang</name><name sortKey="Wang, Chen Ran" sort="Wang, Chen Ran" uniqKey="Wang C" first="Chen Ran" last="Wang">Chen Ran Wang</name><name sortKey="Wong, Freda Pui Fan" sort="Wong, Freda Pui Fan" uniqKey="Wong F" first="Freda Pui-Fan" last="Wong">Freda Pui-Fan Wong</name><name sortKey="Yu, Albert Cheung Hoi" sort="Yu, Albert Cheung Hoi" uniqKey="Yu A" first="Albert Cheung-Hoi" last="Yu">Albert Cheung-Hoi Yu</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Lau, Lok Ting" sort="Lau, Lok Ting" uniqKey="Lau L" first="Lok Ting" last="Lau">Lok Ting Lau</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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