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Pre-PCR processing - Strategies to generate PCR-compatible samples

Rådström, Peter LU ; Knutsson, Rickard LU ; Wolffs, Petra LU ; Lövenklev, Maria LU and Löfström, Charlotta LU (2004) In Molecular Biotechnology 26(2). p.133-146
Abstract
Polymerase chain reaction (PCR) is recognized as a rapid, sensitive, and specific molecular diagnostic tool for the analysis of nucleic acids. However, the sensitivity and kinetics of diagnostic PCR may be dramatically reduced when applied directly to biological samples, such as blood and feces, owing to PCR-inhibitory components. As a result, pre-PCR processing procedures have been developed to remove or reduce the effects of PCR inhibitors. Pre-PCR processing comprises all steps prior to the detection of PCR products, that is, sampling, sample preparation, and deoxyribonucleic acid (DNA) amplification. The aim of pre-PCR processing is to convert a complex biological sample with its target nucleic acids/cells into PCR-amplifiable samples... (More)
Polymerase chain reaction (PCR) is recognized as a rapid, sensitive, and specific molecular diagnostic tool for the analysis of nucleic acids. However, the sensitivity and kinetics of diagnostic PCR may be dramatically reduced when applied directly to biological samples, such as blood and feces, owing to PCR-inhibitory components. As a result, pre-PCR processing procedures have been developed to remove or reduce the effects of PCR inhibitors. Pre-PCR processing comprises all steps prior to the detection of PCR products, that is, sampling, sample preparation, and deoxyribonucleic acid (DNA) amplification. The aim of pre-PCR processing is to convert a complex biological sample with its target nucleic acids/cells into PCR-amplifiable samples by combining sample preparation and amplification conditions. Several different pre-PCR processing strategies are used: (1) optimization of the DNA amplification conditions by the use of alternative DNA polymerases and/or amplification facilitators, (2) optimization of the sample preparation method, (3) optimization of the sampling method, and (4) combinations of the different strategies. This review describes different pre-PCR processing strategies to circumvent PCR inhibition to allow accurate and precise DNA amplification. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
amplification facilitators, PCR-amplifiable samples, PCR-compatible samples, PCR facilitators, PCR inhibitors, PCR sample, pre-PCR processing, sample preparation, thermostable DNA polymerase
in
Molecular Biotechnology
volume
26
issue
2
pages
133 - 146
publisher
Humana Press
external identifiers
  • pmid:14764939
  • wos:000189294400006
  • scopus:4344599509
ISSN
1559-0305
DOI
10.1385/MB:26:2:133
language
English
LU publication?
yes
id
acd2e036-2c56-410d-a3b1-7de8a2f0ed04 (old id 140503)
date added to LUP
2007-06-28 17:06:04
date last changed
2017-12-10 04:39:00
@article{acd2e036-2c56-410d-a3b1-7de8a2f0ed04,
  abstract     = {Polymerase chain reaction (PCR) is recognized as a rapid, sensitive, and specific molecular diagnostic tool for the analysis of nucleic acids. However, the sensitivity and kinetics of diagnostic PCR may be dramatically reduced when applied directly to biological samples, such as blood and feces, owing to PCR-inhibitory components. As a result, pre-PCR processing procedures have been developed to remove or reduce the effects of PCR inhibitors. Pre-PCR processing comprises all steps prior to the detection of PCR products, that is, sampling, sample preparation, and deoxyribonucleic acid (DNA) amplification. The aim of pre-PCR processing is to convert a complex biological sample with its target nucleic acids/cells into PCR-amplifiable samples by combining sample preparation and amplification conditions. Several different pre-PCR processing strategies are used: (1) optimization of the DNA amplification conditions by the use of alternative DNA polymerases and/or amplification facilitators, (2) optimization of the sample preparation method, (3) optimization of the sampling method, and (4) combinations of the different strategies. This review describes different pre-PCR processing strategies to circumvent PCR inhibition to allow accurate and precise DNA amplification.},
  author       = {Rådström, Peter and Knutsson, Rickard and Wolffs, Petra and Lövenklev, Maria and Löfström, Charlotta},
  issn         = {1559-0305},
  keyword      = {amplification facilitators,PCR-amplifiable samples,PCR-compatible samples,PCR facilitators,PCR inhibitors,PCR sample,pre-PCR processing,sample preparation,thermostable DNA polymerase},
  language     = {eng},
  number       = {2},
  pages        = {133--146},
  publisher    = {Humana Press},
  series       = {Molecular Biotechnology},
  title        = {Pre-PCR processing - Strategies to generate PCR-compatible samples},
  url          = {http://dx.doi.org/10.1385/MB:26:2:133},
  volume       = {26},
  year         = {2004},
}