A novel fluorescence quantification method for polymerase chain reaction system

Due to the increasing demand of simultaneous DNA detection, known as multiplexing detection, in a single sample, the current real-time polymerase chain reaction (PCR) platforms are equipped with discrete optics and detectors for different fluorescence wavelength. However, to go beyond four DNA labeling dyes, the optical loss will lower the performance on fluorescence detection. The proposed fluorescence detection system, composing of an ultra-sensitive spectrometer, can provide continuous wavelength detection and can be employed for multiple DNA quantification. The results show that this prototype provides comparable sensitivity and amplification efficiency as Roche Light-Cycler for DNA quantification and similar reproducibility within five intra assay samples with the DNA quantification method proposed in this study.