Abstract
In recent years, the development of polymerase chain reaction (PCR) technology has focused on digital PCR, which depends on the microfluidics. Based on continuous-flow microfluidic technology, this paper designed a miniaturized digital PCR amplification system, and greatly reduced the area required for microdroplet generation and reaction. The core rod. made of polydimethylsiloxane (PDMS), was combined with the Teflon tube to form 3D microfluidics, which requires only one heating source to form the temperature difference required for gene amplification. Only two 34 g needles can form and transmit micro-droplets in a 4-fold tapered Teflon tube, which is the simplest method to generate digital PCR droplets as far as we know, which allows the microdroplet generation device to be free from dependence on expensive chips. A complementary metal oxide semiconductor (CMOS) camera was used as a detection tool to obtain fluorescence video for the entire loop area or a specified loop area. In addition, we developed a homebrew for automatic image acquisition and processing to realize the function of digital PCR. This technique realizes the analysis of clinical serum samples of hepatitis B virus (HBV) and obtained the same results as real-time quantitative PCR. This system has greatly reduced the size and cost of the entire system, while maintaining a stable response.
Highlights
With the development of life science and medical testing, it has become increasingly important that faster and more accurate disease detection devices can be used for more people
At the same time, when their eyes look at the display for a long time, it will cause visual fatigue., which will affect the accuracy of the statistical results
The mainstream development direction of new Polymerase chain reaction (PCR) technology is still microfluidic based on chip PCR
Summary
With the development of life science and medical testing, it has become increasingly important that faster and more accurate disease detection devices can be used for more people. Among the rapidly developing microanalysis systems, micro total analysis systems (μTAS), first proposed by Manz and Widmer in Switzerland in 1990 [1], are the most vigorous. In μTAS, microfluidic chip systems, known as lab-on-a-chip, are at the forefront of current development and have the broadest prospects for development. Polymerase chain reaction (PCR), a method for the enzymatic synthesis of specific DNA in vitro, has been proposed for over 30 years. PCR has developed into the conventional key technology in the field of molecular biology, which has greatly promoted the development of various fields of life sciences. The PCR reaction consists of denaturation, annealing, and extension for one cycle
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