Fluorescence detection test by black printed circuit board based microfluidic channel for polymerase chain reaction

Hwang, Ji-Soo; Kim, Yu-Seop; Song, Hye-Jeong; Kim, Jong-Dae; Park, Chan-Young

doi:10.3233/thc-151061

Cited by 8 publications

(6 citation statements)

References 16 publications

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“…In a previous study, a micro-PCR chip fabricated with double-sided tape produced results similar to those of conventional PCRs [ 9 ]. It was also confirmed that real-time PCR could be implemented using the micro-PCR chip based on a black PCB, as suggested in previous studies [ 10 , 11 ]. Although the heating pattern circuit, which acts as a heater, is sufficiently wide to involve the microfluidic channel, the DNA amplification is successful.…”

Section: Introductionsupporting

confidence: 84%

Performance evaluation of optimal real-time polymerase chain reaction achieved with reduced voltage

Hwang¹,

Kim²,

Kim³

et al. 2018

BioMed Eng OnLine

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BackgroundPolymerase chain reaction (PCR) is used in nucleic acid tests of infectious diseases in point-of-care testing. Previous studies have demonstrated real-time PCR that uses a micro-PCR chip made of packing tape, double-sided tape, and a plastic cover with polycarbonate or polypropylene on a black matte printed circuit board substrate. Despite the success of DNA amplification and fluorescence detection using an early version of the micro-PCR chip, reaching the target temperature was fairly slow and, as a result, the total running time was getting longer. To reduce this runtime, the micro-PCR chip was modified by reducing the heater pattern size of the PCB substrate to one-quarter of the original size or less, while maintaining the ability of the heating pattern to cover the reservoir area of the microfluidic channel. In subsequent experiments, DNA amplification failed several times. During the analysis of the cause of this failure, it was found that the reagent was boiling with the heating range from 25 to 95 °C.MethodsAs a method of DNA amplification verification, images were captured by digital single-lens reflex camera to detect FAM fluorescence using diagonal illumination from a blue LED light source. The images were automatically captured at 72 °C (the extension step in nucleic acid amplification) and the brightness of the captured images was analyzed to con-firm the success of DNA amplification.ResultsCompared to the previous chip with a larger heating pattern size, the current chip appears to generate excess energy as the size of the heating pattern was reduced. To reduce this excess energy, the initial voltage was lowered to 2 V and 2.5 V, which is equivalent to a one-fifth and one-quarter voltage–power reduction in pulse width modulation control, respectively. In both voltage reduction cases, the DNA amplification was successful.ConclusionsDNA amplification tests may fail due to the excess energy generated by reducing the heater pattern size of the PCB substrate. However, the tests succeeded when the voltage was reduced to 2 V or 2.5 V. The 2.5 V power test was more efficient for reducing the overall running time.

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Section: Introductionsupporting

confidence: 84%

Performance evaluation of optimal real-time polymerase chain reaction achieved with reduced voltage

Hwang¹,

Kim²,

Kim³

et al. 2018

BioMed Eng OnLine

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“…Because it was difficult to set this resolution with the standard timer provided by MS Windows, we set it to less than 5 ms using a high-precision event timer (HPET). The advantage of this host-local architecture is a reduction in the overall cost [ 29 , 30 , 31 ].…”

Section: The Proposed Smartphone Camera-based Rt-pcr Systemmentioning

confidence: 99%

“…Generally, the PCR step consists of denaturing DNA or RNA at 94 °C–95 °C, annealing the primer at 58 °C–60 °C, and elongating the DNA or RNA using primers at 72 °C–73 °C. The functions related to the execution and file management of the set PCR step file are processed by the host through the file input/output process in the graphical user interface environment [ 29 , 30 , 31 ].…”

Section: The Proposed Smartphone Camera-based Rt-pcr Systemmentioning

confidence: 99%

“…The PCB substrate was finished in matte black to prevent light reflection, and the thermal spreader was coated with tin to make it easier to distinguish changes in fluorescent brightness [ 33 ]. In our previous version, the thermal spreader was silk-printed with white ink; however, the tin coating delivers a better signal-to-noise ratio than silk [ 29 , 31 ].…”

Section: Pcr Chipmentioning

confidence: 99%

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Quantitative Analysis of Fluorescence Detection Using a Smartphone Camera for a PCR Chip

Kim

Park

Kim

et al. 2021

Sensors

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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.

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“…Real-time PCR technology enables the real-time monitoring of the amplification process for each cycle with a low contamination rate. The procedure also rapidly produces the analyzed results using the fluorescent detection technique [ 7 , 8 ]. With these advantages, many studies have been conducted on the technique and various real-time PCR devices are available.…”

Section: Introductionmentioning

confidence: 99%

Evaluation-independent system for DNA section amplification

Lee

Kim

et al. 2018

BioMed Eng OnLine

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BackgroundIn general, the image analysis of nucleic acid for detecting DNA is dependent on the gel documentation system. These experiments may deal with harmful staining agents and are time consuming. To address these issues, real-time polymerase chain reaction (PCR) devices have been developed. The advantages of real-time PCR are its capabilities for real-time diagnosis, improved sensitivity, and digitization of measurement results. However, real-time PCR equipment is still too bulky and expensive for use in small hospitals and laboratories.MethodsThis paper describes an evaluation-independent real-time PCR system that differs from conventional systems in that it uses a side-illumination optical detection system and a temperature adjustment coefficient for DNA detection. The overall configuration of the evaluation-independent system includes the PCR chip and system hardware and software. The use of the side-illumination method for detection enables the system size to be reduced compared to systems using a typical illumination method. Furthermore, the results of a PCR test are strongly affected by the reaction temperature. Thus, extremely precise control of the temperature of the reaction is needed to obtain accurate results and good reliability. We derived a temperature compensation coefficient that allows us to compensate for the differences between the measured temperature of the negative temperature coefficient (NTC) thermistor sensor and the real temperature of the thermocouple.ResultsApplying the temperature compensation coefficient parameter using the NTC thermistor and using the side-illumination method resulted in an increase in the initial sensor value. The occurrence of the DNA section amplification decreased to 22 cycles from 24 cycles.ConclusionsThe proposed system showed comparable performance to that of an existing real-time PCR, even with the use of simpler and smaller optical devices.

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Fluorescence detection test by black printed circuit board based microfluidic channel for polymerase chain reaction

Cited by 8 publications

References 16 publications

Performance evaluation of optimal real-time polymerase chain reaction achieved with reduced voltage

Performance evaluation of optimal real-time polymerase chain reaction achieved with reduced voltage

Quantitative Analysis of Fluorescence Detection Using a Smartphone Camera for a PCR Chip

Evaluation-independent system for DNA section amplification

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