Hansen Marcus Celik, Nederby Line, Henriksen Mads Okkels-Birk, Hansen Maria, Nyvold Charlotte Guldborg
Biotechniques. 2014 May;56(5):217-28. doi: 10.2144/000114164.
Quantitative PCR (qPCR) of reverse-transcribed mRNA has revolutionized gene expression analyses. qPCR analysis is based on the prevalent assumption that mRNA transcript numbers provide an adequate measure of specific biomarker expression. However, taking the complexity of protein turnover into account, there is a need to correlate qPCR-derived transcriptional patterns with protein translational patterns so as to not leave behind important pathobiological details. One emerging approach in protein analysis is PCR-coupled protein quantification, often denoted as immuno-PCR (iPCR), which targets soluble proteins. Here we review recent trends and applications in iPCR assays that may bridge the gap between classical enzyme-linked immunosorbent assays and mass spectrometry methodologies in terms of sensitivity and multiplexing.
逆转录mRNA的定量PCR（qPCR）彻底改变了基因表达分析。qPCR分析基于一个普遍的假设，即mRNA转录本数量能够充分衡量特定生物标志物的表达。然而，考虑到蛋白质周转的复杂性，有必要将qPCR得出的转录模式与蛋白质翻译模式相关联，以免遗漏重要的病理生物学细节。蛋白质分析中一种新兴的方法是PCR耦合蛋白质定量，通常称为免疫PCR（iPCR），其针对的是可溶性蛋白质。在此，我们综述了iPCR检测的最新趋势和应用，这些趋势和应用在灵敏度和多重分析方面可能弥合经典酶联免疫吸附测定和质谱方法之间的差距。
