Abstract
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.
Highlights
The image analysis of nucleic acid for detecting deoxyribonucleic acid (DNA) is dependent on the gel documentation system
Polymerase chain reaction (PCR) is an important technology used in biological research because it enables disease diagnoses with only a small amount of deoxyribonucleic acid (DNA) [1,2,3]
The polymer‐ ase chain reaction (PCR) system hardware receives the resistance value of the temperature sensor in the PCR chip from the analog-to-digital converter (ADC) and converts it into a temperature value which is transferred to the software via a universal serial bus (USB)
Summary
The image analysis of nucleic acid for detecting DNA is dependent on the gel documentation system. These experiments may deal with harm‐ ful staining agents and are time consuming. To address these issues, real-time polymer‐ ase chain reaction (PCR) devices have been developed. Real-time PCR equipment is still too bulky and expen‐ sive for use in small hospitals and laboratories. The procedure 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. The methods, results, discussion, and conclusions are presented
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