Scalable Transcriptional Analysis Routine—Multiplexed Quantitative Real-Time Polymerase Chain Reaction Platform for Gene Expression Analysis and Molecular Diagnostics
Identifieur interne : 000F93 ( Main/Exploration ); précédent : 000F92; suivant : 000F94Scalable Transcriptional Analysis Routine—Multiplexed Quantitative Real-Time Polymerase Chain Reaction Platform for Gene Expression Analysis and Molecular Diagnostics
Auteurs : Elizabeth P. Garcia ; Lori A. Dowding ; Lawrence W. Stanton ; Vladimir I. SlepnevSource :
- The Journal of molecular diagnostics : JMD [ 1525-1578 ] ; 2005.
Descripteurs français
- KwdFr :
- ARN viral (analyse), ARN viral (génétique), Analyse de profil d'expression de gènes (), Analyse de profil d'expression de gènes (instrumentation), Animaux, Colorants fluorescents, Couleur, Encéphale (métabolisme), Facteurs temps, RT-PCR (), RT-PCR (instrumentation), Rats, Reproductibilité des résultats, Sensibilité et spécificité, Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (virologie), TAQ polymerase (métabolisme), Transcription génétique (génétique), Virus du SRAS (génétique), Virus du SRAS (isolement et purification).
- MESH :
- analyse : ARN viral.
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : ARN viral, Transcription génétique, Virus du SRAS.
- isolement et purification : Virus du SRAS.
- métabolisme : Analyse de profil d'expression de gènes, Encéphale, RT-PCR, TAQ polymerase.
- virologie : Syndrome respiratoire aigu sévère.
- Analyse de profil d'expression de gènes, Animaux, Colorants fluorescents, Couleur, Facteurs temps, RT-PCR, Rats, Reproductibilité des résultats, Sensibilité et spécificité.
English descriptors
- KwdEn :
- Animals, Brain (metabolism), Color, Fluorescent Dyes, Gene Expression Profiling (instrumentation), Gene Expression Profiling (methods), RNA, Viral (analysis), RNA, Viral (genetics), Rats, 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, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (virology), Taq Polymerase (metabolism), Time Factors, Transcription, Genetic (genetics).
- MESH :
- chemical , analysis : RNA, Viral.
- chemical , genetics : RNA, Viral.
- chemical , metabolism : Taq Polymerase.
- chemical : Fluorescent Dyes.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus, Transcription, Genetic.
- instrumentation : Gene Expression Profiling, Reverse Transcriptase Polymerase Chain Reaction.
- isolation & purification : SARS Virus.
- metabolism : Brain.
- methods : Gene Expression Profiling, Reverse Transcriptase Polymerase Chain Reaction.
- virology : Severe Acute Respiratory Syndrome.
- Animals, Color, Rats, Reproducibility of Results, Sensitivity and Specificity, Time Factors.
Abstract
We report the development of a new technology for simultaneous quantitative detection of multiple targets in a single sample. Scalable transcriptional analysis routine (STAR) represents a novel integration of reverse transcriptase-polymerase chain reaction and capillary electrophoresis that allows detection of dozens of gene transcripts in a multiplexed format using amplicon size as an identifier for each target. STAR demonstrated similar or better sensitivity and precision compared to two commonly used methods, SYBR Green-based and TaqMan probe-based real-time reverse transcriptase-polymerase chain reaction. STAR can be used as a flexible platform for building a variety of applications to monitor gene expression, from single gene assays to assays analyzing the expression level of multiple genes. Using severe acute respiratory syndrome (SARS) corona virus as a model system, STAR technology detected single copies of the viral genome in a two-gene multiplex. Blinded studies using RNA extracted from various tissues of a SARS-infected individual showed that STAR correctly identified all samples containing SARS virus and yielded negative results for non-SARS control samples. Using alternate priming strategies, STAR technology can be adapted to transcriptional profiling studies without requiring
Url:
PubMed: 16237214
PubMed Central: 1888488
Affiliations:
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Le document en format XML
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Scalable transcriptional analysis routine (STAR) represents a novel integration of reverse transcriptase-polymerase chain reaction and capillary electrophoresis that allows detection of dozens of gene transcripts in a multiplexed format using amplicon size as an identifier for each target. STAR demonstrated similar or better sensitivity and precision compared to two commonly used methods, SYBR Green-based and TaqMan probe-based real-time reverse transcriptase-polymerase chain reaction. STAR can be used as a flexible platform for building a variety of applications to monitor gene expression, from single gene assays to assays analyzing the expression level of multiple genes. Using severe acute respiratory syndrome (SARS) corona virus as a model system, STAR technology detected single copies of the viral genome in a two-gene multiplex. Blinded studies using RNA extracted from various tissues of a SARS-infected individual showed that STAR correctly identified all samples containing SARS virus and yielded negative results for non-SARS control samples. Using alternate priming strategies, STAR technology can be adapted to transcriptional profiling studies without requiring <italic>a priori</italic> sequence information. Thus, STAR technology offers a flexible platform for development of highly multiplexed assays in gene expression analysis and molecular diagnostics.</p></div></front></TEI><affiliations><list></list><tree><noCountry><name sortKey="Dowding, Lori A" sort="Dowding, Lori A" uniqKey="Dowding L" first="Lori A." last="Dowding">Lori A. Dowding</name><name sortKey="Garcia, Elizabeth P" sort="Garcia, Elizabeth P" uniqKey="Garcia E" first="Elizabeth P." last="Garcia">Elizabeth P. Garcia</name><name sortKey="Slepnev, Vladimir I" sort="Slepnev, Vladimir I" uniqKey="Slepnev V" first="Vladimir I." last="Slepnev">Vladimir I. Slepnev</name><name sortKey="Stanton, Lawrence W" sort="Stanton, Lawrence W" uniqKey="Stanton L" first="Lawrence W." last="Stanton">Lawrence W. Stanton</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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