| Real-time, fluorescence-based quantitative PCR: a snapshot of current procedures and preferences. | |
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MedLine Citation:
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PMID: 16013967 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Real-time fluorescence-based quantitative PCR has become established as the benchmark technology for the quantification of nucleic acids, offering an immense choice of protocols, chemistries and instruments. However, whilst there are comparatively few technical problems associated with DNA-targeted quantitative PCR, this is not the case for real-time reverse transcription PCR assays, and there is considerable uncertainty regarding biological or clinical relevance of many real-time reverse transcription PCR results. A survey of working practices of nearly 100 delegates carried out prior to the Third qPCR Symposium held in London, UK, April 25-26, 2005, reveals some of the reasons underlying the variability of reported real-time reverse transcription PCR results. Specifically, the survey reveals extensive interlaboratory variation in assay design, validation and analysis that, together with other dubious practices, are the likely cause for the publication of variable results. These results emphasize the urgent need for the establishment of best practice guidelines for this technology, particularly in the context of its mounting adaptation as a high-throughput clinical diagnostic assay. |
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Authors:
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Stephen A Bustin |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Expert review of molecular diagnostics Volume: 5 ISSN: 1744-8352 ISO Abbreviation: Expert Rev. Mol. Diagn. Publication Date: 2005 Jul |
Date Detail:
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Created Date: 2005-07-14 Completed Date: 2007-01-25 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101120777 Medline TA: Expert Rev Mol Diagn Country: England |
Other Details:
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Languages: eng Pagination: 493-8 Citation Subset: IM |
Affiliation:
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The Royal London Hospital, Centre for Academic Surgery, 4th Floor Alexandra Wing, London, E1 1BB, UK. s.a.bustin@qmul.ac.uk |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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DNA Primers
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genetics Fluorescence RNA, Messenger / analysis, genetics Research Design Reverse Transcriptase Polymerase Chain Reaction / methods* |
| Chemical | |
Reg. No./Substance:
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0/DNA Primers; 0/RNA, Messenger |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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