One-tube nested RT-PCR enabled by using a plastic film and its application for the rapid detection of SARS-virus.
Identifieur interne : 000722 ( Ncbi/Curation ); précédent : 000721; suivant : 000723One-tube nested RT-PCR enabled by using a plastic film and its application for the rapid detection of SARS-virus.
Auteurs : Sheng-Ce Tao [République populaire de Chine] ; Di Jiang ; Hong-Li Lu ; Wan-Li Xing ; Yu-Xiang Zhou ; Jing ChengSource :
- Biotechnology letters [ 0141-5492 ] ; 2004.
Descripteurs français
- KwdFr :
- ARN viral (analyse), ARN viral (génétique), Analyse de panne d'appareillage, Conception d'appareillage, Matériaux biocompatibles, Polyéthylènes, RT-PCR (), RT-PCR (instrumentation), Reproductibilité des résultats, Sensibilité et spécificité, Virus du SRAS (génétique), Virus du SRAS (isolement et purification).
- MESH :
- analyse : ARN viral.
- génétique : ARN viral, Virus du SRAS.
- isolement et purification : Virus du SRAS.
- Analyse de panne d'appareillage, Conception d'appareillage, Matériaux biocompatibles, Polyéthylènes, RT-PCR, Reproductibilité des résultats, Sensibilité et spécificité.
English descriptors
- KwdEn :
- Biocompatible Materials, Equipment Design, Equipment Failure Analysis, Polyethylenes, RNA, Viral (analysis), RNA, Viral (genetics), Reproducibility of Results, Reverse Transcriptase Polymerase Chain Reaction (instrumentation), Reverse Transcriptase Polymerase Chain Reaction (methods), SARS Virus (genetics), SARS Virus (isolation & purification), Sensitivity and Specificity.
- MESH :
- chemical , analysis : RNA, Viral.
- chemical , genetics : RNA, Viral.
- chemical : Biocompatible Materials, Polyethylenes.
- genetics : SARS Virus.
- instrumentation : Reverse Transcriptase Polymerase Chain Reaction.
- isolation & purification : SARS Virus.
- methods : Reverse Transcriptase Polymerase Chain Reaction.
- Equipment Design, Equipment Failure Analysis, Reproducibility of Results, Sensitivity and Specificity.
Abstract
A general strategy based on the use of a plastic film that allows reverse transcription and nested-PCR in a single closed-tube has been developed. The reaction mixture for the second PCR amplification is quarantined in the cap of the reaction tube during the first round amplification by a piece of plastic film, and later introduced into the PCR amplicons from the first round reaction by centrifugation without opening the reaction tube. The main advantages of our method are its high sensitivity, specificity, simplicity, cost effectiveness, low risk of contamination and the ease in establishment of conditions for nested-PCR. The method has been successfully applied to the detection of the genomic RNA from SARS-CoV, the detection limit of this method is comparable to that of the conventional two-tube nested PCR system. This method could be easily adapted to the detection of other targets.
DOI: 10.1023/b:bile.0000013708.65032.f1
PubMed: 15049359
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pubmed:15049359Le document en format XML
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last="Lu">Hong-Li Lu</name></author><author><name sortKey="Xing, Wan Li" sort="Xing, Wan Li" uniqKey="Xing W" first="Wan-Li" last="Xing">Wan-Li Xing</name></author><author><name sortKey="Zhou, Yu Xiang" sort="Zhou, Yu Xiang" uniqKey="Zhou Y" first="Yu-Xiang" last="Zhou">Yu-Xiang Zhou</name></author><author><name sortKey="Cheng, Jing" sort="Cheng, Jing" uniqKey="Cheng J" first="Jing" last="Cheng">Jing Cheng</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2004">2004</date><idno type="RBID">pubmed:15049359</idno><idno type="pmid">15049359</idno><idno type="doi">10.1023/b:bile.0000013708.65032.f1</idno><idno type="wicri:Area/PubMed/Corpus">002E76</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002E76</idno><idno type="wicri:Area/PubMed/Curation">002E76</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002E76</idno><idno 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type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Jiang, Di" sort="Jiang, Di" uniqKey="Jiang D" first="Di" last="Jiang">Di Jiang</name></author><author><name sortKey="Lu, Hong Li" sort="Lu, Hong Li" uniqKey="Lu H" first="Hong-Li" last="Lu">Hong-Li Lu</name></author><author><name sortKey="Xing, Wan Li" sort="Xing, Wan Li" uniqKey="Xing W" first="Wan-Li" last="Xing">Wan-Li Xing</name></author><author><name sortKey="Zhou, Yu Xiang" sort="Zhou, Yu Xiang" uniqKey="Zhou Y" first="Yu-Xiang" last="Zhou">Yu-Xiang Zhou</name></author><author><name sortKey="Cheng, Jing" sort="Cheng, Jing" uniqKey="Cheng J" first="Jing" last="Cheng">Jing Cheng</name></author></analytic><series><title level="j">Biotechnology letters</title><idno type="ISSN">0141-5492</idno><imprint><date when="2004" type="published">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Biocompatible Materials</term><term>Equipment Design</term><term>Equipment Failure Analysis</term><term>Polyethylenes</term><term>RNA, Viral (analysis)</term><term>RNA, Viral (genetics)</term><term>Reproducibility of Results</term><term>Reverse Transcriptase Polymerase Chain Reaction (instrumentation)</term><term>Reverse Transcriptase Polymerase Chain Reaction (methods)</term><term>SARS Virus (genetics)</term><term>SARS Virus (isolation & purification)</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN viral (analyse)</term><term>ARN viral (génétique)</term><term>Analyse de panne d'appareillage</term><term>Conception d'appareillage</term><term>Matériaux biocompatibles</term><term>Polyéthylènes</term><term>RT-PCR ()</term><term>RT-PCR (instrumentation)</term><term>Reproductibilité des résultats</term><term>Sensibilité et spécificité</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (isolement et purification)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" 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" type="chemical" xml:lang="en"><term>Biocompatible Materials</term><term>Polyethylenes</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>ARN viral</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="instrumentation" xml:lang="en"><term>Reverse Transcriptase Polymerase Chain Reaction</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" xml:lang="en"><term>Equipment Design</term><term>Equipment Failure Analysis</term><term>Reproducibility of Results</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Analyse de panne d'appareillage</term><term>Conception d'appareillage</term><term>Matériaux biocompatibles</term><term>Polyéthylènes</term><term>RT-PCR</term><term>Reproductibilité des résultats</term><term>Sensibilité et spécificité</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A general strategy based on the use of a plastic film that allows reverse transcription and nested-PCR in a single closed-tube has been developed. The reaction mixture for the second PCR amplification is quarantined in the cap of the reaction tube during the first round amplification by a piece of plastic film, and later introduced into the PCR amplicons from the first round reaction by centrifugation without opening the reaction tube. The main advantages of our method are its high sensitivity, specificity, simplicity, cost effectiveness, low risk of contamination and the ease in establishment of conditions for nested-PCR. The method has been successfully applied to the detection of the genomic RNA from SARS-CoV, the detection limit of this method is comparable to that of the conventional two-tube nested PCR system. This method could be easily adapted to the detection of other targets.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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