Nucleic Acid Test Kit Hybridization Solution
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Solution Overview
Problem
Existing reverse hybridization technologies for nucleic acid detection are complex, time-consuming, and costly, especially when detecting multiple samples, often requiring expensive fluorescence detection devices and involving numerous reaction solutions and steps.
Innovation Solution
A test kit with a hybridization solution containing a non-ionic surfactant, a cationic polymer, and a buffer solution with a pH range of 6.5 to 8.5, which enables electrostatic adsorption of biotin-labeled target nucleic acids, preventing non-specific adsorption of alkaline phosphatase and maintaining its activity, allowing for a one-step reaction without the need for separate ELISA steps or extensive washing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional reverse hybridization technology is used for nucleic acid detection, then detection accuracy can be achieved, but the operation process becomes complex and time-consuming with numerous reaction solutions and steps
Solution Approach 1:
The patent combines the hybridization reaction and ELISA detection into a single integrated reaction system. The biotin-labeled target nucleic acid hybridizes with the probe on the solid support, and alkaline phosphatase-labeled streptavidin is added directly to the hybridization solution for one-step detection, eliminating separate washing and binding steps.
Solution Approach 2:
The hybridization solution serves multiple functions simultaneously: it provides the hybridization environment for nucleic acid binding, contains the alkaline phosphatase-labeled streptavidin for detection, and includes components that prevent non-specific adsorption. This multi-functional solution reduces the number of separate reagents and steps.
2Measurement precision
If conventional reverse hybridization technology is used for multiple sample detection, then detection accuracy is maintained, but the detection time is remarkably prolonged
Solution Approach 1:
By merging the hybridization and detection steps into one simultaneous process, the patent eliminates the sequential time requirements of separate operations. Multiple samples can be processed in parallel using the same integrated protocol, significantly reducing total detection time.
Solution Approach 2:
The integrated system allows continuous detection without interrupting for washing or separate binding steps. The alkaline phosphatase-labeled streptavidin remains in solution throughout the hybridization process, enabling continuous monitoring and rapid results.
3Measurement precision
If conventional reverse hybridization technology is used, then detection can be performed, but expensive fluorescence detection devices are required
Solution Approach 1:
The patent uses a colorimetric detection system with alkaline phosphatase and substrates like BCIP/NBT that produce visible color changes. This replaces expensive fluorescence detectors with simple visual or spectrophotometric reading, making the system accessible to routine laboratories.
Solution Approach 2:
The patent replaces the optical fluorescence detection system with a chemical colorimetric system. The alkaline phosphatase enzyme catalyzes substrate conversion producing a color change that can be detected by simple visual inspection or basic spectrophotometry, substituting complex optical equipment with chemical detection.
4Measurement precision
If conventional reverse hybridization technology is used, then detection can be performed, but numerous reaction solutions and extensive washing steps are required
Solution Approach 1:
The patent merges the hybridization buffer and detection reagent into a single solution system. The alkaline phosphatase-labeled streptavidin is added directly to the hybridization solution, eliminating the need for separate washing buffers and reducing overall reagent consumption.
Solution Approach 2:
The cationic polymer acts as an intermediary that prevents non-specific adsorption of alkaline phosphatase to the solid support surface. This mediation allows the enzyme to remain in solution and function effectively without requiring extensive washing to remove adsorbed enzyme, reducing both reagent waste and操作步骤.
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 enables rapid, cost-effective, and high-throughput detection of target nucleic acids with improved specificity and reduced experimental errors, combining nucleic acid hybridization and ELISA into a single step, thereby simplifying operations and reducing reagent consumption.
Implementation Method 1
the hybridization solution contains a non-ionic surfactant, a cationic polymer, and a buffer solution having a pH value in the range from 6.5 to 8.5... enables electrostatic adsorption of biotin-labeled target nucleic acids
Data Source
AI summary
Related to is the field of nucleic acid testing, and in particular, a test kit or a method for testing a target nucleic acid in a sample. The test kit comprises therein a hybridization solution, which contains therein a non-ionic surfactant, a cationic polymer, and a buffer solution having a pH value in the range from 6.5 to 8.5. The test kit can further comprise therein a Tris-HCl color developing solution having a pH value in the range from 9.0 to 10.0 and containing a C8-C18 alkylglucoside. Testing target nucleic acid in a sample using the test kit has the advantages of short time consumption, easy operation, high throughput, and low costs.