Digital nucleic acid amplification detection method and integrated detection system based on CRISPR and Cas

Abstract

The invention disclosed a digital nucleic acid amplification detection method and integrated detection system based on CRISPR / CAS.The integrated detection system includes an integrated reaction chip, temperature control module, light source, and optical signal detectors; the nucleic acid amplification system is divided into amplify microfit droplets, and then the amplifier micro -droplets and containing digital nucleic acid amplification and contains includeThere is a CRISPR / CAS system to detect a mix of micro -droplets to perform a CRISPR reaction. After the reaction is over, the high -specific detection of the target is detected by detecting the optical signal, and the concentration or copy number of the nucleic acid molecules in the sample to be tested to achieve the target of the targetHigh -sensitivity is absolutely quantitative test.The present invention has the characteristics of absolute quantitative quantitative quantitative digital amplification technology, but also the advantages of CRISPR / CAS high -sensitive and high -specific detection. The testing process is integrated on a chip, simplifying the operation steps, and avoiding problems such as cross -pollution.

Description

technical field [0001] The invention relates to the technical field of nucleic acid detection microfluidics, in particular to a CRISPR / Cas-based digital nucleic acid amplification detection method and an integrated detection system. Background technique [0002] Since Vogelstein et al. proposed the digital Polymerase Chain Reaction (dPCR) in 1999, it has shown great technical advantages and application prospects in food safety, forensic identification, precision medicine and other research fields. dPCR is an absolute quantification technique for nucleic acid molecules. By dispersing a sample into tens to tens of thousands of different reaction units, the number of nucleic acid templates in each unit is less than or equal to 1; each unit is separately amplified by PCR After the amplification, the reaction units with nucleic acid templates will emit fluorescent signals, and the reaction units without templates will have no fluorescent signals. Therefore, the number of nucleic ...

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to provide a digital nucleic acid amplification detection method based on CRISPR / Cas and an integrated detection system in order to solve the problems of poor integration of the digital detection system in the background technology and the use of the CRISPR / Cas system to easily cause aerosol pollution. Using this method, CRISPR / Cas is combined with digital nucleic acid amplification technology to achieve absolute quantitative and highly specific detection of target nucleic acid molecules. Integrated detection system for CRISPR / Cas

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