Lee Young-Jo, Kim Daeyoung, Lee Kihoon, Chun Jong-Yoon
Seegene, Inc., Taewon Bldg., 91 Ogeum-ro, Songpa-Gu, Seoul 138-828, South Korea.
Sci Rep. 2014 Dec 11;4:7439. doi: 10.1038/srep07439.
Multiplex real-time PCR with quantification of targets in a single fluorescence channel has been the demand in biotechnology industry. Here, we develop a novel analytical real-time PCR technique to detect multiple targets in a single fluorescence channel without melting curve analysis. In this technique, we show the intensity of the fluorescence signals of two discrete Tm targets is different at certain temperatures called detection temperatures, by which a high Tm target can be detected regardless of a low Tm target. We then identify the low Tm target by utilizing a change of the fluorescence signals between two different detection temperatures. Furthermore, it enables us to determine quantification of each target in a single channel, possibly facilitating convenient patient care for drug treatment in clinics.
在单一荧光通道中对目标进行定量的多重实时PCR一直是生物技术行业的需求。在此，我们开发了一种新型的分析实时PCR技术，无需熔解曲线分析即可在单一荧光通道中检测多个目标。在该技术中，我们发现在称为检测温度的特定温度下，两个离散的解链温度（Tm）目标的荧光信号强度不同，借此可检测高Tm目标而不受低Tm目标的影响。然后，我们利用两个不同检测温度之间荧光信号的变化来识别低Tm目标。此外，它使我们能够在单一通道中确定每个目标的定量，这可能有助于在临床中为药物治疗提供便捷的患者护理。
