Quantitation of severe acute respiratory syndrome coronavirus genome by real-time polymerase chain reaction assay using minor groove binder DNA probe technology.
Identifieur interne : 005178 ( Main/Curation ); précédent : 005177; suivant : 005179Quantitation of severe acute respiratory syndrome coronavirus genome by real-time polymerase chain reaction assay using minor groove binder DNA probe technology.
Auteurs : Hsi-Hsun Lin [République populaire de Chine] ; Shiow-Jen Wang ; Yung-Ching Liu ; Susan Shin-Jung Lee ; Chun-Kai Hwang ; Yao-Shen Chen ; Shue-Ren Wann ; Yi-Li ShihSource :
- Journal of microbiology, immunology, and infection = Wei mian yu gan ran za zhi [ 1684-1182 ] ; 2004.
Descripteurs français
- KwdFr :
- ADN complémentaire (génétique), ADN viral (génétique), Flambées de maladies, Génome viral, Humains, RT-PCR (), Sensibilité et spécificité, Sondes d'ADN (génétique), Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (virologie), Syndrome respiratoire aigu sévère (épidémiologie), Séquence nucléotidique, Taïwan (épidémiologie), Techniques de sonde moléculaire, Virus du SRAS (génétique), Virus du SRAS (isolement et purification).
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : ADN complémentaire, ADN viral, Sondes d'ADN, Virus du SRAS.
- isolement et purification : Virus du SRAS.
- virologie : Syndrome respiratoire aigu sévère.
- épidémiologie : Syndrome respiratoire aigu sévère, Taïwan.
- Flambées de maladies, Génome viral, Humains, RT-PCR, Sensibilité et spécificité, Séquence nucléotidique, Techniques de sonde moléculaire.
- Wicri :
- geographic : Taïwan.
English descriptors
- KwdEn :
- Base Sequence, DNA Probes (genetics), DNA, Complementary (genetics), DNA, Viral (genetics), Disease Outbreaks, Genome, Viral, Humans, Molecular Probe Techniques, 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 (epidemiology), Severe Acute Respiratory Syndrome (virology), Taiwan (epidemiology).
- MESH :
- chemical , genetics : DNA Probes, DNA, Complementary, DNA, Viral.
- geographic , epidemiology : Taiwan.
- diagnosis : Severe Acute Respiratory Syndrome.
- epidemiology : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- isolation & purification : SARS Virus.
- methods : Reverse Transcriptase Polymerase Chain Reaction.
- statistics & numerical data : Reverse Transcriptase Polymerase Chain Reaction.
- virology : Severe Acute Respiratory Syndrome.
- Base Sequence, Disease Outbreaks, Genome, Viral, Humans, Molecular Probe Techniques, Sensitivity and Specificity.
Abstract
The ability to rapidly recognize severe acute respiratory syndrome coronavirus (SARS-CoV) as a cause of infections is critical to quickly limiting further spread of the disease. A rapid, sensitive, and specific laboratory diagnostic test is needed to confirm outbreaks of SARS-CoV infection and distinguish it from other diseases that can cause similar clinical symptoms. An improved TaqMan technology using minor groove binder (MGB) probes was used to detect and quantify SARS-CoV in suspected patients. SARS-CoV primers and probe were designed based on the open reading frame 1b sequence, which encodes coronavirus replicase protein. A linear standard curve with R2 > 0.99 was obtained, and the threshold sensitivity was 10 genome equivalents per reaction. Interassay coefficients of variation were 1.73 to 2.72%, indicating good reproducibility of this method. A total of 228 specimens from 151 suspected patients were quantified by this method, 13 specimens from 6 patients were positive with viral load range from 362 to 36,240,000 genome equivalents/mL. In conclusion, the high sensitivity and reproducibility of the real-time polymerase chain reaction SARS-CoV RNA quantitation using MGB probe allowed the screening of large numbers of clinical samples.
PubMed: 15497005
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technology.</title><author><name sortKey="Lin, Hsi Hsun" sort="Lin, Hsi Hsun" uniqKey="Lin H" first="Hsi-Hsun" last="Lin">Hsi-Hsun Lin</name><affiliation wicri:level="1"><nlm:affiliation>Section of Infectious Diseases, Department of Medicine, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan, ROC.</nlm:affiliation><country xml:lang="fr" wicri:curation="lc">République populaire de Chine</country><wicri:regionArea>Section of Infectious Diseases, Department of Medicine, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan</wicri:regionArea><wicri:noRegion>Taiwan</wicri:noRegion></affiliation></author><author><name sortKey="Wang, Shiow Jen" sort="Wang, Shiow Jen" uniqKey="Wang S" first="Shiow-Jen" last="Wang">Shiow-Jen Wang</name></author><author><name sortKey="Liu, Yung Ching" sort="Liu, Yung Ching" uniqKey="Liu Y" first="Yung-Ching" last="Liu">Yung-Ching Liu</name></author><author><name sortKey="Lee, Susan Shin Jung" sort="Lee, Susan Shin Jung" uniqKey="Lee S" first="Susan Shin-Jung" last="Lee">Susan Shin-Jung Lee</name></author><author><name sortKey="Hwang, Chun Kai" sort="Hwang, Chun Kai" uniqKey="Hwang C" first="Chun-Kai" last="Hwang">Chun-Kai Hwang</name></author><author><name sortKey="Chen, Yao Shen" sort="Chen, Yao Shen" uniqKey="Chen Y" first="Yao-Shen" last="Chen">Yao-Shen Chen</name></author><author><name sortKey="Wann, Shue Ren" sort="Wann, Shue Ren" uniqKey="Wann S" first="Shue-Ren" last="Wann">Shue-Ren Wann</name></author><author><name sortKey="Shih, Yi Li" sort="Shih, Yi Li" uniqKey="Shih Y" first="Yi-Li" last="Shih">Yi-Li Shih</name></author></analytic><series><title level="j">Journal of microbiology, immunology, and infection = Wei mian yu gan ran za zhi</title><idno type="ISSN">1684-1182</idno><imprint><date when="2004" type="published">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Base Sequence</term><term>DNA Probes (genetics)</term><term>DNA, 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respiratoire aigu sévère (diagnostic)</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Syndrome respiratoire aigu sévère (épidémiologie)</term><term>Séquence nucléotidique</term><term>Taïwan (épidémiologie)</term><term>Techniques de sonde moléculaire</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (isolement et purification)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA Probes</term><term>DNA, Complementary</term><term>DNA, Viral</term></keywords><keywords scheme="MESH" type="geographic" qualifier="epidemiology" xml:lang="en"><term>Taiwan</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="epidemiology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN complémentaire</term><term>ADN viral</term><term>Sondes d'ADN</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></keywords><keywords scheme="MESH" qualifier="statistics & numerical data" xml:lang="en"><term>Reverse Transcriptase 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" qualifier="épidémiologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term><term>Taïwan</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term><term>Disease Outbreaks</term><term>Genome, Viral</term><term>Humans</term><term>Molecular Probe Techniques</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Flambées de maladies</term><term>Génome viral</term><term>Humains</term><term>RT-PCR</term><term>Sensibilité et spécificité</term><term>Séquence nucléotidique</term><term>Techniques de sonde moléculaire</term></keywords><keywords scheme="Wicri" type="geographic" xml:lang="fr"><term>Taïwan</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The ability to rapidly recognize severe acute respiratory syndrome coronavirus (SARS-CoV) as a cause of infections is critical to quickly limiting further spread of the disease. A rapid, sensitive, and specific laboratory diagnostic test is needed to confirm outbreaks of SARS-CoV infection and distinguish it from other diseases that can cause similar clinical symptoms. An improved TaqMan technology using minor groove binder (MGB) probes was used to detect and quantify SARS-CoV in suspected patients. SARS-CoV primers and probe were designed based on the open reading frame 1b sequence, which encodes coronavirus replicase protein. A linear standard curve with R2 > 0.99 was obtained, and the threshold sensitivity was 10 genome equivalents per reaction. Interassay coefficients of variation were 1.73 to 2.72%, indicating good reproducibility of this method. A total of 228 specimens from 151 suspected patients were quantified by this method, 13 specimens from 6 patients were positive with viral load range from 362 to 36,240,000 genome equivalents/mL. In conclusion, the high sensitivity and reproducibility of the real-time polymerase chain reaction SARS-CoV RNA quantitation using MGB probe allowed the screening of large numbers of clinical samples.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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