Pre-PCR processing |
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| Authors: | Peter Rådström Rickard Knutsson Petra Wolffs Maria Lövenklev Charlotta Löfström |
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| Institution: | (1) Applied Microbiology, Lund Institute of Technology, Lund University, PO Box 124, SE-221 00 Lund, Sweden |
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| 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. |
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| Keywords: | Amplification facilitators PCR-amplifiable samples PCR-compatible samples PCR facilitators PCR inhibitors PCR sample pre-PCR processing sample preparation thermostable DNA polymerase |
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