Nucleic Acid Biosensor Using Primer Bubble Regions for Multiplexed Detection
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Solution Overview
Problem
Current methods for detecting biomolecules, such as nucleotide sequences, face challenges in accuracy and efficiency due to limitations in traditional symptom-based diagnoses and the need for improved detection techniques that can handle multiple analytes simultaneously with high sensitivity and specificity.
Innovation Solution
A method involving the amplification of target nucleotide sequences using primers with a 5' anchor region, a 3' extension region, and a bubble region, followed by nuclease cleavage to generate probes immobilized on conducting nanostructures, allowing for simultaneous detection of presence, level, and sequence variations through changes in electrical charge.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional symptom-based diagnosis methods are used, then the diagnostic process is simple, but the accuracy and reliability of diagnosis deteriorates
Solution Approach 1:
The detection method is segmented into distinct functional modules: nucleic acid amplification module, probe hybridization module, and electrical signal detection module. Each module performs a specific function, allowing the complex detection process to be broken down into manageable, optimized components that collectively achieve high accuracy without overwhelming complexity
Solution Approach 2:
Probe molecules serve as intermediaries between the target nucleic acid sequences and the electrical detection system. These probes hybridize specifically to target sequences and translate molecular recognition events into measurable electrical signals through changes in charge distribution, enabling accurate detection while maintaining a clear separation between biological and electrical domains
2Productivity
If traditional single-analyte detection methods are used, then the detection process is straightforward, but the productivity and efficiency deteriorates due to inability to handle multiple analytes simultaneously
Solution Approach 1:
The detection system is designed with universal components that can detect multiple different analytes simultaneously. The same electrical detection mechanism and signal processing architecture handle various nucleic acid targets by simply changing the probe sequences, enabling high-throughput multiplexed detection without requiring separate specialized systems for each analyte
Solution Approach 2:
The system exploits changes in electrical charge parameters to differentiate between multiple analytes. Different probe-target hybridization events produce distinct charge change signatures that can be simultaneously measured and resolved, allowing multiplexed detection through parameter differentiation rather than requiring physically separate detection channels
3Measurement precision
If amplification of target nucleotide sequence is performed, then the sensitivity of detection is improved, but the time required for detection increases
Solution Approach 1:
Nucleic acid amplification is performed as a preliminary action before the actual detection step. By pre-amplifying the target sequences to abundant levels, the subsequent hybridization and detection steps can proceed rapidly with high sensitivity, separating the time-consuming amplification phase from the quick detection phase to minimize total detection time
Solution Approach 2:
The system replaces traditional mechanical or optical detection methods with electrical field-based detection. This substitution enables rapid signal acquisition immediately after hybridization without requiring lengthy washing, drying, or optical scanning steps, significantly reducing the time component while maintaining high sensitivity through the amplification step
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
Enables sensitive and specific detection of nucleotide sequences, facilitating accurate diagnosis and multiplexing capabilities, thereby improving the reliability of biomolecular detection and analysis.
Implementation Method 1
contacting the analyte with the probe under hybridizing conditions
Implementation Method 2
the signal detected by the foregoing method can be generated when hybridization of the analyte to the probe causes a change in the electrical charge of the probe
Implementation Method 3
the one or more primers may comprise nucleotide sequences that are cleaved by a nuclease, wherein the nuclease may be a restriction enzyme of DNA, and/or RNA
Data Source
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AI summary
Embodiments of the present invention relate generally to strategies and methods of amplifying short target sequences using one or more primers comprising a 5' anchor region, a 3' extension region, and a bubble region therebetween.