Rådström Peter, Knutsson Rickard, Wolffs Petra, Lövenklev Maria, Löfström Charlotta
Applied Microbiology, Lund Institute of Technology, Lund University, PO Box 124, SE-221 00 Lund, Sweden.
Mol Biotechnol. 2004 Feb;26(2):133-46. doi: 10.1385/MB:26:2:133.
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.
聚合酶链反应（PCR）是一种用于核酸分析的快速、灵敏且特异的分子诊断工具。然而，由于存在PCR抑制成分，当直接应用于生物样本（如血液和粪便）时，诊断性PCR的灵敏度和动力学可能会显著降低。因此，已开发出PCR前处理程序以去除或减少PCR抑制剂的影响。PCR前处理包括检测PCR产物之前的所有步骤，即采样、样本制备和脱氧核糖核酸（DNA）扩增。PCR前处理的目的是通过结合样本制备和扩增条件，将含有目标核酸/细胞的复杂生物样本转化为可进行PCR扩增的样本。使用了几种不同的PCR前处理策略：（1）通过使用替代DNA聚合酶和/或扩增促进剂优化DNA扩增条件，（2）优化样本制备方法，（3）优化采样方法，以及（4）不同策略的组合。本综述描述了不同的PCR前处理策略，以规避PCR抑制，从而实现准确且精确的DNA扩增。
