Development of two Novel PCR Machines base on the Capillary Convective Polymerase Chain Reaction
Date Issued
2011
Date
2011
Author(s)
Chang, Wei-Chen
Abstract
In this study, we developed two novel prototypes of platform base on the capillary convective polymerase chain reaction（CCPCR）technology, including onsite detection
type and real-time detection type. CCPCR is a novel technology for polymerase chain reaction. It only needs one single temperature controller under the bottom of glass capillary tube in which the sample is contained, and the bottom temperature is maintained at about 95℃. Thus, the temperature gradient between the bottom and the top will drive a thermal convection loop that can successfully execute PCR within 30 minutes.
For the onsite detection type platform, we applied the positive temperature coefficient thermistor (PTC Thermistor) as the bottom heating components. PTC
thermistor provides a flat surface and a steady temperature control for the CCPCR due to its special self-heating characteristic. Overall, the advantages of the detection
device are compact, simple, low cost, portable, low power consumption, powered by a lithium battery, and perform the high sensitivity of nucleic acid detection to make
disease detection more quickly and easily.
In addition, for the real-time CCPCR platform design, we set up the excitation light source on the bottom of the glass capillary and the sensor on the top for fluorescence detection. The optical system and the accurate temperature control will be able to quantitatively identify the initial concentration of the template DNA. Four samples can be simultaneously execute CCPCR. For the quantitative analysis, we define a Ct (Crossing time) value. According to the results, the logarithm of the initial DNA
concentration and the Ct value shows linear correlation which can be taken as a calibration curve in real-time CCPCR. Compared to the commercial real-time PCR platform, the cost of ours platform is quite low.
Based on the results of this study, we can promote this technology for the family and personal care after doing the improved design to become optimized, commercialized and humanized. We believe that these platforms will have a great contribution in medical detection in the future.
Subjects
Convection
Polymerase chain reaction
Nucleic acid detection
Portable
Real-time nucleic acid detection
SDGs
Type
thesis
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