Rådström Peter, Löfström Charlotta, Lövenklev Maria, Knutsson Rickard, Wolffs Petra
CSH Protoc. 2008 Mar 1;2008:pdb.top20. doi: 10.1101/pdb.top20.
INTRODUCTIONThe use of conventional and real-time PCR is to some extent restricted by the presence of PCR inhibitors. This is particularly so when the techniques are applied directly to complex biological samples such as clinical, environmental, or food samples for the detection of microorganisms. PCR inhibitors can originate from the sample itself, or as a result of the method used to collect or otherwise prepare the sample. Either way, inhibitors can dramatically reduce the sensitivity and amplification efficiency of PCR. This article discusses methods of reducing inhibition and designing reliable and sensitive conventional and real-time PCR experiments.
引言
常规PCR和实时PCR的应用在一定程度上受到PCR抑制剂的限制。当这些技术直接应用于复杂的生物样本(如临床、环境或食品样本)以检测微生物时,情况尤其如此。PCR抑制剂可能源于样本本身,或者是由于用于收集或以其他方式制备样本的方法。无论哪种方式,抑制剂都能显著降低PCR的灵敏度和扩增效率。本文讨论了减少抑制作用以及设计可靠且灵敏的常规PCR和实时PCR实验的方法。