Skrzypski Marcin
Medical University of Gdansk, Department of Allergology, 7 Debinki St, 80-211 Gdansk, Poland.
Lung Cancer. 2008 Feb;59(2):147-54. doi: 10.1016/j.lungcan.2007.11.008. Epub 2008 Jan 4.
Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) is rapidly becoming a basic method in lung cancer research. Analysis of transcriptional activity of tumor cells or detection of tumor markers by this technique has the potential to change lung cancer diagnosis and treatment. Quantitative RT-PCR is characterized by unparalleled sensitivity and specificity, with very reliable reproducibility. Its prime advantage for gene expression analysis is its broad dynamic range of 10(7)-fold. Moreover, it is cost-effective, feasible in every day laboratory routine and efficient in terms of biological material consumption. Still, there are a number of methodological aspects that need to be carefully considered before it can sensibly be implemented into clinical practice. Three major technical issues: the choice of chemistries, gene expression data normalization and statistical processing of the results will be specifically highlighted in this review. Further, clinical applications of qRT-PCR will be thoroughly discussed: detection and staging of lung cancer and construction and validation of prognostic and predictive gene expression signatures.
定量逆转录聚合酶链反应（qRT-PCR）正迅速成为肺癌研究中的一种基本方法。通过该技术分析肿瘤细胞的转录活性或检测肿瘤标志物，有可能改变肺癌的诊断和治疗方式。定量RT-PCR具有无与伦比的灵敏度和特异性，重复性非常可靠。其在基因表达分析方面的主要优势是具有10（7）倍的广泛动态范围。此外，它具有成本效益，在日常实验室常规操作中可行，并且在生物材料消耗方面效率很高。然而，在将其合理应用于临床实践之前，仍有许多方法学方面的问题需要仔细考虑。本综述将特别强调三个主要技术问题：化学方法的选择、基因表达数据的标准化以及结果的统计处理。此外，还将深入讨论qRT-PCR的临床应用：肺癌的检测和分期以及预后和预测性基因表达特征的构建与验证。
