Valasek Mark A, Repa Joyce J
Department of Physiology Touchstone Center for Diabetes Research, University of Texas Southwestern Medical Center, Dallas, Texas 75390-8854, USA.
Adv Physiol Educ. 2005 Sep;29(3):151-9. doi: 10.1152/advan.00019.2005.
In recent years, real-time polymerase chain reaction (PCR) has emerged as a robust and widely used methodology for biological investigation because it can detect and quantify very small amounts of specific nucleic acid sequences. As a research tool, a major application of this technology is the rapid and accurate assessment of changes in gene expression as a result of physiology, pathophysiology, or development. This method can be applied to model systems to measure responses to experimental stimuli and to gain insight into potential changes in protein level and function. Thus physiology can be correlated with molecular events to gain a better understanding of biological processes. For clinical molecular diagnostics, real-time PCR can be used to measure viral or bacterial loads or evaluate cancer status. Here, we discuss the basic concepts, chemistries, and instrumentation of real-time PCR and include present applications and future perspectives for this technology in biomedical sciences and in life science education.
近年来,实时聚合酶链反应(PCR)已成为一种强大且广泛应用于生物学研究的方法,因为它能够检测和定量极少量的特定核酸序列。作为一种研究工具,该技术的一个主要应用是快速、准确地评估由于生理、病理生理或发育而导致的基因表达变化。此方法可应用于模型系统,以测量对实验刺激的反应,并深入了解蛋白质水平和功能的潜在变化。因此,生理学可以与分子事件相关联,从而更好地理解生物过程。对于临床分子诊断,实时PCR可用于测量病毒或细菌载量或评估癌症状态。在此,我们讨论实时PCR的基本概念、化学原理和仪器设备,并介绍该技术在生物医学科学和生命科学教育中的当前应用及未来前景。