Amplified Nucleic Acid Detection via Sandwich Hybridization
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Solution Overview
Problem
Current nucleic acid detection methods for genetic diagnosis are costly, time-consuming, and require specialized equipment, making them burdensome for patients and testers, and they struggle with the simultaneous detection of multiple targets and the sensitivity of single-stranded DNA detection.
Innovation Solution
A method involving the amplification of double-stranded nucleic acids with single-stranded regions at each end, which are hybridized with oligonucleotide probes on a solid phase, allowing for visual detection without specialized equipment, using a device with chromatographic carriers and labeled probes for enhanced sensitivity and cost-effectiveness.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescent labeling is used for nucleic acid detection, then detection sensitivity is improved, but cost increases significantly
Solution Approach 1:
The patent replaces expensive fluorescent labels with inexpensive colloidal gold particles as detection markers. The colloidal gold provides visual detection capability without the high cost of fluorescent labeling, achieving a balance between detection sensitivity and cost-effectiveness through the use of cheap alternative materials.
Solution Approach 2:
The patent utilizes the inherent color properties of colloidal gold particles for visual detection. The color change or color presence indicates detection results, eliminating the need for expensive fluorescent labels while maintaining detectability through simple visual observation rather than requiring complex fluorescent detection equipment.
2Stability of the object's composition
If heating treatment is applied for denaturation, then double-stranded nucleic acid is separated into single strands, but hybridization efficiency decreases due to reannealing
Solution Approach 1:
The patent changes the denaturation method from thermal heating to chemical denaturation using alkaline solutions. This parameter change in the denaturation approach allows for effective strand separation without the reannealing problem caused by heating, thereby maintaining high hybridization efficiency while achieving complete denaturation.
Solution Approach 2:
The patent introduces alkaline solutions as an intermediary substance to facilitate denaturation. Instead of using heat directly, the alkaline environment acts as a mediator to separate the double-stranded nucleic acid into single strands, preventing reannealing and preserving hybridization efficiency for subsequent detection steps.
3Adaptability or versatility
If multiple target nucleic acids are detected simultaneously using antigen-antibody pairs, then detection capability is improved, but the number of available combinations is limited
Solution Approach 1:
The patent implements a universal detection system where a single colloidal gold-labeled probe can detect multiple different target nucleic acids. By designing the probe to recognize conserved regions or using a panel of probes with the same detection mechanism, the system achieves multiplex detection capability without being constrained by the limited number of antigen-antibody pairs, thereby expanding versatility while maintaining simplicity.
4Ease of operation
If single-stranded DNA is used for detection, then hybridization is simplified, but detection sensitivity decreases due to curling into balls
Solution Approach 1:
The patent introduces double-stranded nucleic acid structures as intermediaries in the detection process. Rather than directly detecting single-stranded DNA that curls into balls, the system uses double-stranded structures that maintain stable configurations and can be effectively detected, thereby preserving detection sensitivity while simplifying the overall hybridization procedure through the use of these stable intermediate forms.
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
This approach enables rapid, accurate, and cost-effective detection of amplified DNA fragments, allowing for simultaneous analysis of multiple targets without the need for expensive labels or specialized equipment, improving detection sensitivity and reducing procedural complexity.
Implementation Method 1
hybridizing a first oligonucleotide probe immobilized on a solid phase with one of single-stranded regions
Implementation Method 2
transferred a liquid sample containing the randomly sampled genes or fragments thereof by capillary action
Data Source
AI summary
An object of the invention is to provide a nucleic acid detection method which takes advantage of the high specificity of hybridization techniques, reduces the time length and the number of steps required for detection of PCR products, and allows for easy and highly accurate detection by visual observation without the need of special equipment; and a nucleic acid detection device or kit. The invention provides a method for detecting a target nucleic acid in a sample, which includes performing amplification of the target nucleic acid sequence to synthesize an amplification product having a partially double-stranded structure where a single-stranded region is added to each end of the target sequence, and hybridizing a nucleic acid sequence bound to a development medium and a nucleic acid sequence labeled with a labeling compound with the single-stranded regions of the amplification product to form a sandwich hybridization complex; and a detection device thereof.