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PMID: 11884626 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Review

Real-time PCR in virology.

Nucleic acids research ·Vol. 30 ·No. 6 ·2002-03-15 ·Pages 1292-305

Mackay IM, Arden KE, Nitsche A

Abstract

The use of the polymerase chain reaction (PCR) in molecular diagnostics has increased to the point where it is now accepted as the gold standard for detecting nucleic acids from a number of origins and it has become an essential tool in the research laboratory. Real-time PCR has engendered wider acceptance of the PCR due to its improved rapidity, sensitivity, reproducibility and the reduced risk of carry-over contamination. There are currently five main chemistries used for the detection of PCR product during real-time PCR. These are the DNA binding fluorophores, the 5' endonuclease, adjacent linear and hairpin oligoprobes and the self-fluorescing amplicons, which are described in detail. We also discuss factors that have restricted the development of multiplex real-time PCR as well as the role of real-time PCR in quantitating nucleic acids. Both amplification hardware and the fluorogenic detection chemistries have evolved rapidly as the understanding of real-time PCR has developed and this review aims to update the scientist on the current state of the art. We describe the background, advantages and limitations of real-time PCR and we review the literature as it applies to virus detection in the routine and research laboratory in order to focus on one of the many areas in which the application of real-time PCR has provided significant methodological benefits and improved patient outcomes. However, the technology discussed has been applied to other areas of microbiology as well as studies of gene expression and genetic disease.

MeSH Terms
DNA, Viral/analysis Endonucleases/chemistry Fluorescence Fluorescent Dyes/chemistry Humans Kinetics Oligonucleotide Probes/chemistry Polymerase Chain Reaction/methods Viruses/genetics,isolation & purification
Chemicals
DNA, Viral Fluorescent Dyes Oligonucleotide Probes Endonucleases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Mackay Ian M
Clinical Virology Research Unit, Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Brisbane, Australia. i.mackay@mailbox.uq.edu.au
Arden Katherine E
Nitsche Andreas
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/30.6.1292
Email
i.mackay@mailbox.uq.edu.au
Published
2002-03-15
Pages
1292-305
Language
English
Region
England
NLM ID
0411011
PMCID
PMC101343
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/30.6.1292 PMC Full Text View journal details
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