Nucleic Acid Separation Using Disposable Spin Columns
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Solution Overview
Problem
Current methods for nucleic acid separation, such as the bind-elute method, require skilled personnel and specialized equipment, are costly, and pose infection risks due to the handling of pathogenic organisms, and existing alternatives like zeolite-based methods fail to achieve high-purity nucleic acid concentration and purification.
Innovation Solution
A specimen treatment reagent containing a thiol reductant, alkaline substance, and chaotropic agent is used to release nucleic acids from specimens, which are then adsorbed by a silica carrier, facilitating efficient separation and purification, especially in viscous samples like sputum, while reducing infectivity risks.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If the bind-elute method is used for nucleic acid purification, then high-purity nucleic acids can be obtained, but skilled researchers and specialized equipment are required
Solution Approach 1:
The invention uses disposable spin columns with pre-loaded silica carriers and reagents, eliminating the need for reusable equipment and specialized laboratory infrastructure. Each spin column is a single-use device that integrates all necessary components for nucleic acid purification, making the method accessible to facilities without advanced equipment or trained personnel
Solution Approach 2:
The spin column system is designed to be self-contained and self-operating. The columns automatically perform dissolution, adsorption, washing, and elution steps through simple centrifugation or vacuum pressure, requiring no manual intervention beyond sample loading and reagent addition. This eliminates the need for skilled operators to manage complex procedures
2Manufacturing precision
If the bind-elute method is used for nucleic acid purification, then high-purity nucleic acids can be obtained, but time and cost increase due to outsourcing
Solution Approach 1:
By providing affordable disposable spin columns with integrated reagents, the invention enables facilities to perform nucleic acid purification in-house at low cost, eliminating the need to outsource tests to external institutions and thereby reducing both time and expense
3Measurement precision
If conventional genetic test equipment is used, then nucleic acid analysis can be performed, but infection risk increases due to handling pathogenic organisms
Solution Approach 1:
The spin column system extracts and isolates nucleic acids from pathogenic organisms within a closed microenvironment. The silica carrier selectively binds nucleic acids while leaving infectious agents trapped in the waste stream, which is discarded after a single use. This separation eliminates infection risk while preserving nucleic acid integrity for analysis
Solution Approach 2:
The silica carrier acts as an intermediary that selectively binds nucleic acids from the specimen while leaving pathogenic organisms in the liquid phase. This mediator enables the separation of infectious agents from their genetic material, allowing safe handling of purified nucleic acids without exposure to live pathogens
4Ease of operation
If zeolite-based purification method is used, then simplification is achieved, but nucleic acid concentration and purification quality are insufficient
Solution Approach 1:
The invention changes the adsorption parameters by using silica carriers with optimized surface properties and pore structures, along with controlled chaotropic agent concentrations. This enables high-affinity nucleic acid binding while maintaining procedural simplicity, achieving both ease of operation and high purification quality that zeolite methods cannot attain
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 method enables efficient, high-purity nucleic acid separation and concentration from various specimens, including those with low cell concentrations, reducing the need for advanced equipment and skilled labor, and effectively reduces infection risks through effective disinfection and fluidity improvement.
Implementation Method 1
a component for reducing infectivity of a microorganism, the component for causing a nucleic acid to be released, and a reductant, thereby improving fluidity of the specimen
Implementation Method 2
a specimen treatment reagent for causing a nucleic acid in a specimen to be released
Implementation Method 3
allowing a nucleic acid released from the specimen to be adsorbed by a carrier comprising silica
Data Source
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AI summary
An object of the present invention is to provide a method for efficiently separating a nucleic acid from a specimen containing a nucleic acid without the use of specific equipment, and a device therefor. The method of the present invention uses a device comprising a treatment reagent-accommodating container 100 accommodating a treatment reagent for causing a nucleic acid to be released 101 and a nucleic acid-collecting member 200 comprising a nucleic acid adsorptive carrier 205, which is configured such that a cover 104 is attached to the treatment reagent-accommodating container 100, and when the treatment reagent-accommodating container 100 is connected to the nucleic acid-collecting member 200, a seal with the cover 104 is opened, thereby allowing a discharge opening 102 and a treatment reagent-supplying opening 204 to be communicated with each other, the method comprising: step 1 of accommodating a specimen in the nucleic acid-collecting member 200; step 2 of connecting the treatment reagent-accommodating container 100 to the nucleic acid-collecting member 200, thereby causing the treatment reagent-accommodating container 100 to be communicated with the nucleic acid-collecting member 200; step 3 of mixing the specimen and the treatment reagent to obtain a mixture 421, thereby causing a nucleic acid to be released; and step 4 of allowing the mixture 421 to pass through the carrier 205 so as to be discharged, thereby allowing the carrier 205 to adsorb the nucleic acid.