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1996年美国Applied Biosystems公司推出了实时荧光定量PCR技术,该技术是指在PCR反应体系中加入荧光基团,利用荧光信号积累实时监测整个PCR进程,最后通过标准曲线对未知模板进行定量分析的方法[1-2]。该技术实现了PCR从定性到定量的飞跃,可动态实时检测核酸产物的形成,无需后处理过程,避免了扩增产物的污染,而且具有直观、重复性好、特异性强、敏感性高、定量准确、自动化程度高、速度快、全封闭和易操作等优点,因而目前已得到广泛应用[3-4]。
Applied Biosystems in the United States in 1996 introduced real-time fluorescence quantitative PCR technology, which refers to the PCR reaction system by adding fluorophores, the use of fluorescence signal accumulation in real-time monitoring of the entire PCR process, and finally through the standard curve of unknown template quantitative analysis method [1-2]. This technology realizes the qualitative and quantitative leap from PCR to real-time detection of the formation of nucleic acid products in real time without the need of post-processing and avoids the pollution of the amplification products. Moreover, the technology has the advantages of being intuitive, good repeatability, high specificity, high sensitivity, Quantitative accuracy, high degree of automation, speed, fully enclosed and easy to operate, etc., which has been widely used [3-4].