Integrated Microdroplet Chip for Low-Concentration Multi-Index PCR
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Solution Overview
Problem
Digital PCR technology faces challenges in low-concentration nucleic acid detection scenarios, particularly in distinguishing multiple rare target sequences due to reduced sensitivity from sample splitting, leading to inaccurate detection results.
Innovation Solution
An integrated droplet chip with one-to-many mixing and reaction structures, including primary and secondary reaction chambers, and a fluorescence detection area, which amplifies samples to higher concentrations before dividing them into droplets for secondary amplification and detection, ensuring sensitivity and accuracy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If the sample is divided into multiple portions for detection, then the number of detection targets is increased, but the sensitivity is reduced
Solution Approach 1:
The detection system is segmented into multiple independent reaction chambers (first reaction chamber, second reaction chamber, third reaction chamber) that can process different targets simultaneously. Each chamber maintains complete sample processing capability while enabling parallel multi-target detection, thus increasing versatility without sacrificing sensitivity through sample division.
Solution Approach 2:
The patent performs preliminary amplification in the first reaction chamber before the droplets are distributed to subsequent reaction chambers. This pre-amplification step ensures that even when samples are divided into multiple portions for different targets, each portion retains sufficient concentration for sensitive detection, thereby resolving the contradiction between multi-target capability and sensitivity.
2Adaptability or versatility
If multiple fluorescent probes are used to detect multiple targets, then the detection capacity is increased, but the spectral distribution limitation is encountered
Solution Approach 1:
The system divides the detection function across multiple spatially separated reaction chambers, each capable of detecting different targets with specific fluorescent probes. This segmentation allows the use of multiple fluorescent probes with different spectral properties without interference, as each probe operates in its own dedicated chamber, thereby increasing detection capacity while overcoming spectral limitations.
3Adaptability or versatility
If the sample concentration is low, then the detection of rare targets becomes possible, but the detection accuracy is reduced
Solution Approach 1:
The first reaction chamber performs preliminary amplification of the low-concentration sample before the amplified product is distributed to subsequent reaction chambers for further processing and detection. This pre-amplification step significantly increases the sample concentration, thereby improving detection accuracy while maintaining the ability to detect rare targets that were present in the original low-concentration sample.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The solution enhances the sensitivity and accuracy of low-concentration nucleic acid detection by increasing sample concentration through pre-amplification, allowing for multi-index detection of rare sequences while integrating primary amplification, mixing, droplet generation, and detection on a single chip, improving integration and automation.
Implementation Method 1
performing primary amplification, so that a detection sample with a first concentration in the primary system is amplified to form a detection sample with a second concentration
Implementation Method 2
a plurality of secondary reaction chambers, which are controllably connected to the plurality of sampling chambers in a one-to-one correspondence, are used to store the droplets generated by the detection sample of the second concentration and the secondary system at the droplet generating structure
Implementation Method 3
a fluorescence detection area which is used for detecting droplets after secondary amplification
Data Source
AI summary
The present application provides an integrated droplet chip and a droplet multi-index detection method therefor and belongs to the technical field of digital PCR analyzers. The chip comprises a chip body, the chip body being constructed with at least one set of one-to-many mixing and reaction structures, a droplet generating structure and a fluorescence detection area, and each of the one-to-many mixing and reaction structures comprises: a primary reaction chamber, used for performing primary amplification, so that a detection sample with a first concentration in the primary system is amplified to form a detection sample with a second concentration, wherein the second concentration is higher than the first concentration; a plurality of sampling chambers; and the plurality of sampling chambers are controllably connected to the same primary reaction chamber at the same time; a plurality of secondary reaction chambers, which are controllably connected to the plurality of sampling chambers in a one-to-one correspondence, are used to store the droplets generated by the detection sample of the second concentration and the secondary system at the droplet generating structure and perform secondary amplification. The present application effectively increases the concentration of the detection sample. The detection sample having the increased concentration is divided into more detection samples to ensure that each detection sample has a relatively high concentration, thereby achieving multi-index detection of a low-concentration sample, ensuring the sensitivity of sample detection, and improving the accuracy of detection results.