Real-time PCR technology for cancer diagnostics.
Real-time PCR technology for cancer diagnostics.
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DOI：
10.1093/clinchem/48.8.1178
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发表时间：
2002-08
影响因子：
9.3
通讯作者：
P. Bernard;C. Wittwer
中科院分区：
文献类型：
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作者：
P. Bernard;C. Wittwer
背景生物科学和技术的进步为诊断和治疗癌症提供了分子靶点。目前外科病理学对恶性肿瘤分期的分类主要基于解剖特征(例如，肿瘤转移)和组织病理学(例如，分级)。微阵列和聚类算法一起揭示了癌症之间的分子多样性，这有望形成一种具有预后意义，更重要的是治疗意义的新分类法。病理学的挑战将是为常规临床实践开发和实施这些分子分类。本文讨论了在临床实验室进行实体肿瘤分析的好处、挑战和可能性，重点介绍了基于DNA的聚合酶链式反应技术。内容分子标记可用于提供准确的预后，并预测对治疗的反应、耐药性或毒性。恶性肿瘤涉及的基因组变化的多样性需要各种分析方法来完成肿瘤的侧写。一些新的肿瘤分子分类是基于基因表达的，需要在样本处理方面进行范式转换，以保持用于分析的RNA的完整性。更稳定的标记物(即DNA和蛋白质)在临床实验室很容易处理。实时定量聚合酶链式反应可以确定基因的重复或缺失。此外，在聚合酶链式反应后立即进行的熔解曲线分析可以识别微小的突变，小到单碱基的变化。这些技术正在变得更容易、更快，并且可以被多路复用。实时定量聚合酶链式反应方法是癌症标志物分析的有利选择。摘要有必要将肿瘤学研究中的最新发现转化为临床实践。这就需要客观、可靠、成本效益高的分子技术进行临床试验，并最终实现常规使用。实时定量聚合酶链式反应在临床实验室肿瘤分析中具有诱人的特点。
BACKGROUND Advances in the biological sciences and technology are providing molecular targets for diagnosing and treating cancer. Current classifications in surgical pathology for staging malignancies are based primarily on anatomic features (e.g., tumor-node-metastasis) and histopathology (e.g., grade). Microarrays together with clustering algorithms are revealing a molecular diversity among cancers that promises to form a new taxonomy with prognostic and, more importantly, therapeutic significance. The challenge for pathology will be the development and implementation of these molecular classifications for routine clinical practice. APPROACH This article discusses the benefits, challenges, and possibilities for solid-tumor profiling in the clinical laboratory with an emphasis on DNA-based PCR techniques. CONTENT Molecular markers can be used to provide accurate prognosis and to predict response, resistance, or toxicity to therapy. The diversity of genomic alterations involved in malignancy necessitates a variety of assays for complete tumor profiling. Some new molecular classifications of tumors are based on gene expression, requiring a paradigm shift in specimen processing to preserve the integrity of RNA for analysis. More stable markers (i.e., DNA and protein) are readily handled in the clinical laboratory. Quantitative real-time PCR can determine gene duplications or deletions. Furthermore, melting curve analysis immediately after PCR can identify small mutations, down to single base changes. These techniques are becoming easier and faster and can be multiplexed. Real-time PCR methods are a favorable option for the analysis of cancer markers. SUMMARY There is a need to translate recent discoveries in oncology research into clinical practice. This requires objective, robust, and cost-effective molecular techniques for clinical trials and, eventually, routine use. Real-time PCR has attractive features for tumor profiling in the clinical laboratory.