Abstract
Most existing commercial real-time polymerase chain reaction (RT-PCR) instruments are bulky because they contain expensive fluorescent detection sensors or complex optical structures. In this paper, we propose an RT-PCR system using a camera module for smartphones that is an ultra small, high-performance and low-cost sensor for fluorescence detection. The proposed system provides stable DNA amplification. A quantitative analysis of fluorescence intensity changes shows the camera’s performance compared with that of commercial instruments. Changes in the performance between the experiments and the sets were also observed based on the threshold cycle values in a commercial RT-PCR system. The overall difference in the measured threshold cycles between the commercial system and the proposed camera was only 0.76 cycles, verifying the performance of the proposed system. The set calibration even reduced the difference to 0.41 cycles, which was less than the experimental variation in the commercial system, and there was no difference in performance.
Highlights
Real-time polymerase chain reaction (RT-PCR), referred to as quantitative polymerase chain reaction, is used to quantitatively measure the amount of amplification of DNA during the PCR process [1,2,3]
The RT-PCR technique has been actively implemented using PCR chips combined with lab-on-a-chip functionality that integrates a variety of technologies such as microfluidic channels [4,5]
We propose a method to optimize the parameter for the threshold cycle, which is an important measure of quantitative analysis, and to compare the threshold cycles obtained with the optimized parameter to those of the reference commercial system
Summary
Real-time polymerase chain reaction (RT-PCR), referred to as quantitative polymerase chain reaction (qPCR), is used to quantitatively measure the amount of amplification of DNA during the PCR process [1,2,3]. PCBs can eliminate most of the obstacles imposed by other boards in the commercialization of microfluidic devices and systems: expansion, standardization and system-level integration of microfluidic devices at a minimal cost These benefits come from the use of well-established PCB technology that is routinely used in the mass production of electronic circuits and consumer electronics [8]. Our team proposed an RT-PCR chip system using a DSLR camera as a fluorescence detection sensor in previous studies [8,9]. We propose a small RT-PCR system that illuminates blue LEDs on a PCR chip diagonally and detects fluorescence using a miniature surveillance camera module with a CMOS sensor for smartphones. Experimental results on three different concentrations of a DNA reagent showed that the difference between the threshold cycles measured by the proposed system and those from the reference commercial system was only 0.41 cycles
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