Real-time PCR in virology

Mackay, I. M., Arden, K. E. and Nitsche, A. (2002) Real-time PCR in virology. Nucleic Acids Research, 30 6: 1292-1305. doi:10.1093/nar/30.6.1292


Author Mackay, I. M.
Arden, K. E.
Nitsche, A.
Title Real-time PCR in virology
Journal name Nucleic Acids Research   Check publisher's open access policy
ISSN 0305-1048
ISBN 0305-1048; 1362-4954; 1362-4962; 0261-3166; 0952-3472
Publication date 2002-03-15
Year available 2005
Sub-type Critical review of research, literature review, critical commentary
DOI 10.1093/nar/30.6.1292
Open Access Status DOI
Volume 30
Issue 6
Start page 1292
End page 1305
Total pages 14
Editor R.T. Walker
D. Soll
A.S. Jones
Place of publication Oxford, U. K.
Publisher Oxford University Press
Language eng
Subject C1
270303 Virology
730101 Infectious diseases
730204 Child health
0601 Biochemistry and Cell Biology
1101 Medical Biochemistry and Metabolomics
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.
Formatted 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.
© 2002 Oxford University Press
Keyword Biochemistry & molecular biology
Polymerase-chain-reaction
Quantitative RT-PCR
Hepatitis-C-virus
Epstein-Barr-virus
Nucleic-acid hybridization
Resonance energy-transfer
Human cytomegalovirus DNA
Varicella-zoster virus
Bone-marrow transplantation
Reverse transcription PCR
Q-Index Code C1
Institutional Status UQ

Document type: Journal Article
Sub-type: Critical review of research, literature review, critical commentary
Collections: Faculty of Science Publications
Excellence in Research Australia (ERA) - Collection
School of Chemistry and Molecular Biosciences
 
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Created: Wed, 15 Aug 2007, 03:07:45 EST