The lack of portability and high cost of multiplex real-time PCR systems limits the device to be used in POC. To overcome this issue, this paper proposes a compact and cost-effective fluorescence detection system that can be integrated to a multiplex real-time PCR equipment. An open platform camera with embedded lens was used instead of photodiodes or an industrial camera. A compact filter wheel using a sliding tape is integrated, and the excitation LEDs are fixed at a 45° angle near the PCR chip, eliminating the need of additional filter wheels. The results show precise positioning of the filter wheel with an error less than 20 μm. Fluorescence detection results using a reference dye and standard DNA amplification showed comparable performance to that of the photodiode system.
This paper proposes a cost-effective real-time multiplexed polymerase chain reaction (PCR) chip system for point-of-care (POC) testing. In the proposed system, nucleic acid amplification is performed in a reaction chamber built on a printed-circuit-board (PCB) substrate with a PCB pattern heater and a thermistor. Fluorescence can be detected through the transparent plastic on the other side of the substrate. Open platform cameras were used for miniaturization and cost-effectiveness. We also used simple and cost-effective oblique lighting to stimulate fluorescence. Response performance was investigated by observing the change in the average brightness of the chamber images with various reference dye concentrations. In addition, we investigated the interference properties between different colors by measuring the fluorescence response for each dye concentration mixed with the maximum concentration of the different dyes. Quantitative performance was validated using standard DNA solutions. Experimental results show that the proposed system is suitable for POC real-time multi-PCR systems.
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