Implementation system of high-throughput integrated micro-droplet digital PCR (Polymerase Chain Reaction)

Abstract

The invention relates to an implementation system of high-throughput integrated micro-droplet digital PCR (Polymerase Chain Reaction). The implementation system comprises a micro-droplet preparation unit, a PCR amplification unit and a fluorescence detection and analysis unit; the micro-droplet preparation unit comprises a driver, a disturbance device and a micro-droplet generator; the micro-droplet generator comprises an oil phase channel, a sample channel and a detection chamber; the driver generates a driving signal and acts on the disturbance device; the disturbance device generates a speed pulse in an oil phase, and then the oil phase shears a mixed liquid of a sample and a PCR reaction reagent to generate micro-droplets; the micro-droplets are uniformly spread in the detection chamber, and the PCR amplification unit performs PCR amplification reaction on the micro-droplets in the detection chamber; and then the fluorescence detection and analysis unit performs data reading and analysis on the micro-droplets to complete quantitative detection of specific nucleic acid. Rapid preparation of the micro-droplets is realized, the oil consumption is reduced, the dependence of the sizes of the micro-droplets on the size of a fluid channel is reduced, the detection precision and reliability are high, and the cost is low.

Description

technical field [0001] The invention relates to a high-throughput integrated micro-droplet digital PCR realization system, which belongs to the technical field of nucleic acid detection and analysis. Background technique [0002] In the field of life science research, PCR (polymerase chain reaction) is an extremely common nucleic acid detection and analysis method. At present, when performing nucleic acid detection, the mainstream droplet digital PCR instrument usually includes three parts: a droplet generation system, a thermal cycler, and a signal reading system. The phase (continuous phase) squeezes the sample water phase (discrete phase) to disperse the sample into several micro-droplets, then place the sample that has become micro-droplets in a sample tube, and place the sample tube in a thermal cycler for amplification. , and then put the sample tube in the signal reading system for detection. The detection principle of the traditional signal reading system is: micro-...

AI Technical Summary

Problems solved by technology

[0003] Traditional micro-droplet generation mostly adopts a passive method, and multiple parameters such as droplet size, frequency and device channel size, flow rate, and physical properties are interrelated, and it is not easy to adjust

Even with fixed operating parameters, droplet size is difficult to precisely control due to batch-to-batch variation in chip size

Passive methods are used to prepare micro-droplets, and the size of the required fluid channel is correspondingly relatively small, requiring high machining accuracy and increasing the cost; if a larger channel size is used, the flow rate of the oil phase must be increased, and redundant fluid must be removed in the follow-up work. Oil, complex process

[0004] At present, in the research of preparing micro-droplets in an active way, most of the external disturbance frequency and droplet generation frequency are not in one-to-one correspondence, and the droplet generation frequency and size still need to be calibrated; and the droplet generation frequency is generally low (< 100Hz); the droplet size range is narrow

[0005] In addition, the current micro-droplet digital PCR involves micro-droplet preparation, amplification reaction, and detection and analysis. At present, related products are completed with multiple equipment and multiple steps. Droplet generation, PCR, and detection are three different systems, and the operation is relatively complicated. There are many factors that affect the final result of PCR, each test takes a long time, and the cost of equipment is high. The tested samples need to be processed separately to prevent cross-contamination

Images