Document Detail


Real-time, fluorescence-based quantitative PCR: a snapshot of current procedures and preferences.
MedLine Citation:
PMID:  16013967     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Stephen A Bustin
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Expert review of molecular diagnostics     Volume:  5     ISSN:  1744-8352     ISO Abbreviation:  Expert Rev. Mol. Diagn.     Publication Date:  2005 Jul 
Date Detail:
Created Date:  2005-07-14     Completed Date:  2007-01-25     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101120777     Medline TA:  Expert Rev Mol Diagn     Country:  England    
Other Details:
Languages:  eng     Pagination:  493-8     Citation Subset:  IM    
Affiliation:
The Royal London Hospital, Centre for Academic Surgery, 4th Floor Alexandra Wing, London, E1 1BB, UK. s.a.bustin@qmul.ac.uk
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MeSH Terms
Descriptor/Qualifier:
DNA Primers / genetics
Fluorescence
RNA, Messenger / analysis,  genetics
Research Design
Reverse Transcriptase Polymerase Chain Reaction / methods*
Chemical
Reg. No./Substance:
0/DNA Primers; 0/RNA, Messenger

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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