Compact PCR Apparatus with Real-Time Fluorescent Signal Detection
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Solution Overview
Problem
Conventional PCR machines are bulky and time-consuming due to repeated heating and cooling, and they require excessive consumables and multiple specimens for multi-target gene quantification, leading to issues with signal crosstalk and inaccurate data analysis.
Innovation Solution
A compact PCR apparatus with a reagent container holding mechanism made from heat-resistant materials, incorporating conductive thin films and temperature sensors, along with a processor for real-time signal analysis, enables rapid heating and cooling, and simultaneous detection of multiple fluorescent signals in a single container using a built-in algorithm.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional PCR machines use repeated heating and cooling cycles, then DNA amplification can be achieved, but the device becomes bulky and reaction time increases
Solution Approach 1:
The patent divides the heating and cooling functions into separate independent modules: a heating unit with heating film and a cooling unit with Peltier elements. This segmentation allows each module to operate independently and simultaneously, eliminating the sequential operation of conventional machines and significantly reducing reaction time while maintaining a compact structure.
Solution Approach 2:
The reaction chamber serves multiple functions: it is both the heating zone and cooling zone, and also the detection chamber for fluorescence signals. This multi-functionality eliminates the need for separate chambers for different operations, reducing device complexity and size while maintaining efficient thermal cycling and real-time detection capabilities.
2Measurement precision
If multiple target genes are quantified using multiple test tubes, then accurate detection is achieved, but consumable usage and specimen requirements increase
Solution Approach 1:
The patent combines multiple detection targets into a single reaction chamber by using spectrally distinct fluorescent dyes for different target genes. The detection unit captures and separates these fluorescent signals based on their unique wavelengths, enabling simultaneous quantification of multiple targets in one test tube, thereby reducing consumable usage and specimen requirements while maintaining detection accuracy.
3Adaptability or versatility
If multiple fluorescent dyes are used for multi-target detection, then detection capability increases, but signal crosstalk occurs
Solution Approach 1:
The patent introduces a detection unit with spectral separation capability as an intermediary between the fluorescent dyes and the detection system. This unit separates the fluorescent signals based on their unique wavelengths using optical filters or spectrometers, effectively eliminating signal crosstalk and enabling accurate simultaneous detection of multiple targets with different fluorescent dyes.
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 apparatus significantly reduces the size and reaction time of PCR processes while accurately quantifying multiple target genes in a single test tube, minimizing consumable usage and specimen requirements, and effectively mitigating signal crosstalk.
Implementation Method 1
The surface layer of the heat-resistant material may include a heat-resistant insulating material or a conductive thin film, which is formed by coating with conductive material
Implementation Method 2
the conductive thin film has specific resistance value, which depends on the design of heating mechanism of the reagent container holding mechanism
Implementation Method 3
the surface layer of the heat-resistant material may include a heat-resistant insulating material or a conductive thin film
Implementation Method 4
The portion, in contact with the reagent container, of the reagent container holding mechanism is provided with at least one temperature sensor
Implementation Method 5
a specific fluorescent probe is added, after each PCR amplification cycle, the DNA of a target gene is amplified and a fluorescent signal is generated simultaneously
Implementation Method 6
when a PCR product is generated, after the specific probe is hydrolyzed, the quencher loses the efficacy in quenching the reporter, so that the fluorescence of the reporter can be detected
Data Source
AI summary
The present invention discloses a Polymerase Chain Reaction (PCR) apparatus for real-time detecting of one or more fluorescent signals. According to the apparatus, the PCR is performed by controlling heating and cooling intervals of a reagent container receiving space. With the aid of an added specific probe and fluorescent material, as well as a light source and a spectrometer, a generated fluorescent signal is detected. Meanwhile, the apparatus is also pre-loaded with an algorithm configured to analyze and quantify the fluorescent signal in a real-time manner.