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

• 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.

Ämnesord

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)

Nyckelord

Amplification facilitators

PCR facilitators

PCR inhibitors

PCR sample

PCR-amplifiable samples

PCR-compatible samples

Pre-PCR processing

Sample preparation

Thermostable DNA polymerase

antibody

betaine

bovine serum albumin

detergent

dimethyl sulfoxide

DNA binding protein

DNA polymerase

glycerol

hemoglobin

immunoglobulin G

lactoferrin

macrogol 400

organic solvent

polymer

polysorbate 20

aqueous solution

bacterial growth

biosynthesis

blood

centrifugation

diagnostic accuracy

diagnostic procedure

dilution

DNA extraction

DNA synthesis inhibition

enrichment culture

feces

filtration

gene amplification

human

nonhuman

nucleic acid analysis

polymerase chain reaction

review

sampling

Animals

Blood Chemical Analysis

DNA

Humans

Microchemistry

Specimen Handling

Bacteria (microorganisms)

Bovinae

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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