Development of a method for concentrating and purifying SARS coronavirus RNA by a magnetic bead capture system.
Identifieur interne : 004867 ( Main/Exploration ); précédent : 004866; suivant : 004868Development of a method for concentrating and purifying SARS coronavirus RNA by a magnetic bead capture system.
Auteurs : Ai-Lian Hei [République populaire de Chine] ; Jian-Ping CaiSource :
- DNA and cell biology [ 1044-5498 ] ; 2005.
Descripteurs français
- KwdFr :
- ARN viral (isolement et purification), Amorces ADN, Diagnostic précoce, Humains, Hybridation d'acides nucléiques, Magnétisme, RT-PCR (), Sensibilité et spécificité, Sondes oligonucléotidiques, Streptavidine, Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (virologie), Virus du SRAS (génétique).
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Virus du SRAS.
- isolement et purification : ARN viral.
- virologie : Syndrome respiratoire aigu sévère.
- Amorces ADN, Diagnostic précoce, Humains, Hybridation d'acides nucléiques, Magnétisme, RT-PCR, Sensibilité et spécificité, Sondes oligonucléotidiques, Streptavidine.
English descriptors
- KwdEn :
- DNA Primers, Early Diagnosis, Humans, Magnetics, Nucleic Acid Hybridization, Oligonucleotide Probes, RNA, Viral (isolation & purification), Reverse Transcriptase Polymerase Chain Reaction (methods), SARS Virus (genetics), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (virology), Streptavidin.
- MESH :
- chemical , isolation & purification : RNA, Viral.
- chemical : DNA Primers, Oligonucleotide Probes, Streptavidin.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- methods : Reverse Transcriptase Polymerase Chain Reaction.
- virology : Severe Acute Respiratory Syndrome.
- Early Diagnosis, Humans, Magnetics, Nucleic Acid Hybridization, Sensitivity and Specificity.
Abstract
Severe acute respiratory syndrome (SARS) is a recently emerged infectious disease caused by a novel coronavirus, which has been designated the SARS coronavirus (SARS-CoV). To date, molecular assays for the detection of SARS-CoV has focused mainly on reverse transcriptase-PCR (RT-PCR) analysis of specimens. However, RT-PCR assays currently available have low sensitivity during the early stage of the disease in which the viral load in specimens is very low. A method for concentrating and purifying SARS-CoV RNA by a magnetic bead capture system was developed and followed by an RT-PCR assay in this study with the goal of improving the sensitivity of the RT-PCR method. This approach takes advantage of the cooperative interaction between adjacently hybridized oligonucleotides. A capture probe was covalently coupled to magnetic beads and a second probe, which anneals adjacent to the capture probe site, was prehybridized in solution to the target. It was shown that, when applied to SARS RNA samples, the sensitivity of nucleic acid capture RTPCR was about 10-fold greater than routine RT-PCR. This nucleic acid capture system was effective in improving the sensitivity of the RT-PCR, due to enriching and purifying SARS-CoV RNA. The method will be helpful for the early detection of the SARS-associated coronavirus.
DOI: 10.1089/dna.2005.24.479
PubMed: 16101344
Affiliations:
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Le document en format XML
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concentrating and purifying SARS coronavirus RNA by a magnetic bead capture system.</title><author><name sortKey="Hei, Ai Lian" sort="Hei, Ai Lian" uniqKey="Hei A" first="Ai-Lian" last="Hei">Ai-Lian Hei</name><affiliation wicri:level="3"><nlm:affiliation>National Center for Clinical Laboratory, Beijing Hospital, Beijing, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Center for Clinical Laboratory, Beijing Hospital, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Cai, Jian Ping" sort="Cai, Jian Ping" uniqKey="Cai J" first="Jian-Ping" last="Cai">Jian-Ping Cai</name></author></analytic><series><title level="j">DNA and cell biology</title><idno type="ISSN">1044-5498</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>DNA Primers</term><term>Early Diagnosis</term><term>Humans</term><term>Magnetics</term><term>Nucleic Acid Hybridization</term><term>Oligonucleotide Probes</term><term>RNA, Viral (isolation & purification)</term><term>Reverse Transcriptase Polymerase Chain Reaction (methods)</term><term>SARS Virus (genetics)</term><term>Sensitivity and Specificity</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term><term>Severe Acute Respiratory Syndrome (virology)</term><term>Streptavidin</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN viral (isolement et purification)</term><term>Amorces ADN</term><term>Diagnostic précoce</term><term>Humains</term><term>Hybridation d'acides nucléiques</term><term>Magnétisme</term><term>RT-PCR ()</term><term>Sensibilité et spécificité</term><term>Sondes oligonucléotidiques</term><term>Streptavidine</term><term>Syndrome respiratoire aigu sévère (diagnostic)</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Virus du SRAS (génétique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>RNA, Viral</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>DNA Primers</term><term>Oligonucleotide Probes</term><term>Streptavidin</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="isolement et purification" xml:lang="fr"><term>ARN viral</term></keywords><keywords scheme="MESH" qualifier="methods" 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" xml:lang="en"><term>Early Diagnosis</term><term>Humans</term><term>Magnetics</term><term>Nucleic Acid Hybridization</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Amorces ADN</term><term>Diagnostic précoce</term><term>Humains</term><term>Hybridation d'acides nucléiques</term><term>Magnétisme</term><term>RT-PCR</term><term>Sensibilité et spécificité</term><term>Sondes oligonucléotidiques</term><term>Streptavidine</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome (SARS) is a recently emerged infectious disease caused by a novel coronavirus, which has been designated the SARS coronavirus (SARS-CoV). To date, molecular assays for the detection of SARS-CoV has focused mainly on reverse transcriptase-PCR (RT-PCR) analysis of specimens. However, RT-PCR assays currently available have low sensitivity during the early stage of the disease in which the viral load in specimens is very low. A method for concentrating and purifying SARS-CoV RNA by a magnetic bead capture system was developed and followed by an RT-PCR assay in this study with the goal of improving the sensitivity of the RT-PCR method. This approach takes advantage of the cooperative interaction between adjacently hybridized oligonucleotides. A capture probe was covalently coupled to magnetic beads and a second probe, which anneals adjacent to the capture probe site, was prehybridized in solution to the target. It was shown that, when applied to SARS RNA samples, the sensitivity of nucleic acid capture RTPCR was about 10-fold greater than routine RT-PCR. This nucleic acid capture system was effective in improving the sensitivity of the RT-PCR, due to enriching and purifying SARS-CoV RNA. The method will be helpful for the early detection of the SARS-associated coronavirus.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><settlement><li>Pékin</li></settlement></list><tree><noCountry><name sortKey="Cai, Jian Ping" sort="Cai, Jian Ping" uniqKey="Cai J" first="Jian-Ping" last="Cai">Jian-Ping Cai</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Hei, Ai Lian" sort="Hei, Ai Lian" uniqKey="Hei A" first="Ai-Lian" last="Hei">Ai-Lian Hei</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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