Strategies to Generate PCR-Compatible Samples
Strategies to Generate PCR-Compatible Samples
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发表时间：
2004
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通讯作者：
P. Rådström;R. Knutsson;P. Wolffs;Maria Lövenklev;C. Löfström
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作者：
P. Rådström;R. Knutsson;P. Wolffs;Maria Lövenklev;C. Löfström
聚合酶链反应（PCR）是公认的用于核酸分析的快速、灵敏和特异的分子诊断工具。然而，当直接应用于生物样品如血液和粪便时，由于PCR抑制组分，诊断PCR的灵敏度和动力学可能显著降低。因此，已经开发了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PCR产物检测前的所有步骤，即取样、样品制备和脱氧核糖核酸（DNA）扩增。PCR前处理的目的是通过组合样品制备和扩增条件将具有其靶核酸/细胞的复杂生物样品转化为PCR可扩增的样品。使用几种不同的PCR前处理策略：（1）通过使用替代的DNA聚合酶和/或扩增促进剂来优化DNA扩增条件，（2）优化样品制备方法，（3）优化取样方法，以及（4）不同策略的组合。本文综述了不同的PCR前处理策略，以规避PCR抑制，使准确和精确的DNA扩增。索引号：扩大促进者; PCR可扩增样本; PCR兼容样本;聚合酶链反应因子; PCR抑制剂; PCR样品; PCR前处理;样品制备;耐热DNA聚合酶。
Pre-PCR Processing 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 dra- matically 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. Index Entries: Amplification facilitators; PCR-amplifiable samples; PCR-compatible samples; PCR fa- cilitators; PCR inhibitors; PCR sample; pre-PCR processing; sample preparation; thermostable DNA poly- merase.