Multilayer three-dimensional digital nucleic acid amplification chip as well as manufacturing and using methods thereof

Abstract

The invention relates to the field of digital PCR, in particular to a multilayer three-dimensional digital nucleic acid amplification chip as well as manufacturing and using methods thereof. According to the multi-layer three-dimensional digital nucleic acid amplification chip, multi-layer superposition of microarray chips can be realized, and the number of reaction units for digital nucleic acid amplification is greatly increased, so that the dynamic range of nucleic acid quantification and the detection lower limit precision are greatly improved, and the reaction flux is greatly increased in actual detection. In addition, the invention further provides the manufacturing method and the using method of the multi-layer three-dimensional digital nucleic acid amplification device, and the accuracy of quantitative results can be further improved.

Description

technical field [0001] The invention relates to the field of digital PCR, in particular to a multilayer three-dimensional digital nucleic acid amplification chip and a method for making and using the chip. Background technique [0002] Since the invention of PCR in 1983, nucleic acid quantification has developed from qualitative technology (electrophoresis after PCR) to semi-quantitative (fluorescence quantitative PCR, qPCR), and with the emergence of digital PCR (digital PCR, dPCR), it has developed to absolute quantification. Digital PCR technology randomly assigns nucleic acid templates to a large number of reaction units for amplification reactions. After the amplification, the fluorescence signal of each reaction unit is collected. Since the random assignment of nucleic acid molecules conforms to the Poisson distribution, the Poisson distribution is finally passed. The original concentration or content of the sample can be obtained by calculating the loose distribution ...

AI Technical Summary

Problems solved by technology

Although some foreign studies have reported digital PCR chips that can generate millions of reaction partitions, they have adopted a strategy of increasing the surface density of microwells on the chip. Although this approach increases the number of reaction units, the volume of the reaction units decreases sharply. , improves the accuracy but cannot take into account the sensitivity, the application range is narrow, and cannot meet the needs of molecular diagnosis such as early cancer screening and new coronavirus diagnosis

Moreover, the increase in surface density leads to an increase in chip manufacturing requirements and a loss rate of finished products, resulting in higher costs.

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