Study On The Self-priming And Discretization Microfluidic Chip For The Digital Nuclear Acid Amplification

By using the digital nucleic acid amplification technology, one can absolutely quantify the number of the target nucleic acid molecule in the sample. Discretization of the target molecule into the individual reaction unit is the first step for the performance of the digital nucleic acid amplification. So, it is meaningful to develop an automatic, handy and efficient platform for the sample discretization of the digital nucleic acid amplification. In this paper, with the help of the soft lithography technology, we have developed a microfluidics chip which can perform the sample self-priming and discretization for the digital nucleic acid amplification. Then, we tested and optimized the chip’s parameters, including the sample injection speed, the uniform of the sample discretization, the evaporation and the heating transfer capacity. Eventually, we successfully performed the digital loop-mediated isothermal amplification (digital-LAMP) and digital PCR on this chip. Taking advantage of power-free sequential sample injection method, it is unnecessary to use any external equipment to drive the sample, and this significantly proved the portability of the microfluidics chip. Employing the so-called sample self-digitization technology can rapidly realize the sample discretization into thousands of individual unit without any complicated valve, and this can reduce the complexity of the chip fabrication. Using LAMP for digital nucleic acid amplification can lower the requirement of the heating equipment for the temperature control. In sum, our sample self-priming and discretization chip for the digital nucleic acid amplification is highly portable, convenient and suitable for the point of care test and rapid assay.