Zeolite Purification for Nucleic Acid Amplification Samples
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Solution Overview
Problem
Current nucleic acid amplification methods face challenges in accurately detecting target nucleic acid from biological samples like expectorate and saliva due to inhibitory substances such as proteins, polysaccharides, and lipids, leading to non-reproducible results and incomplete evaluation of target nucleic acid presence or expression levels.
Innovation Solution
A method involving the use of a nucleic acid extraction reagent with an anionic surfactant and zeolite for purification, where zeolite effectively adsorbs inhibitory substances while allowing target nucleic acid to pass through, resulting in a sample suitable for high-sensitivity nucleic acid amplification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If nucleic acid extraction reagents containing surfactants are used to lyse cells and extract nucleic acid, then nucleic acid can be obtained from biological samples, but surfactants remain in the extract and inhibit nucleic acid amplification reaction
Solution Approach 1:
The patent extracts and removes surfactants from the nucleic acid extract using a specific purification method. The extract is treated with a solution containing divalent metal ions and polyethylene glycol, which selectively binds to and removes surfactants while leaving nucleic acid intact, thus eliminating the harmful surfactant inhibition effect.
Solution Approach 2:
The patent introduces an intermediary purification step using divalent metal ions and polyethylene glycol as mediators. These intermediaries facilitate the selective removal of surfactants from the extract, acting as a bridge between the extraction reagents and the final amplification reaction, thereby resolving the conflict between efficient extraction and clean amplification.
2Ease of operation
If biological samples are directly used for nucleic acid amplification, then the sample processing is simple, but inhibitory substances such as proteins, polysaccharides and lipids partially or completely inhibit the amplification reaction
Solution Approach 1:
The patent performs preliminary purification actions on the biological sample before amplification. The sample undergoes cell lysis, nucleic acid extraction, and surfactant removal treatments in advance, creating a cleaned-up extract that is ready for reliable amplification. This preliminary processing eliminates inhibitory substances that would otherwise compromise reaction reliability.
3Object-affected harmful factors
If the nucleic acid amplification sample is diluted to reduce surfactant concentration, then the amplification reaction can proceed, but the sensitivity and accuracy of detection are reduced
Solution Approach 1:
Instead of diluting the sample to reduce surfactant concentration, the patent extracts and removes surfactants selectively from the extract. This purification approach eliminates the need for dilution, maintaining the original nucleic acid concentration and thus preserving detection sensitivity and accuracy while still achieving surfactant-free conditions suitable for amplification.
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 reproducible detection of target nucleic acid from challenging biological samples without diluting or removing viscosity, improving the accuracy and sensitivity of nucleic acid amplification across various sample types, including blood, spinal fluid, and saliva.
Implementation Method 1
the nucleic acid extract is contacted with zeolite to obtain a nucleic acid amplification sample suitable for nucleic acid amplification
Data Source
AI summary
The object is to remove substances that can inhibit nucleic acid amplification from biological samples and nucleic acid extracts, and to allow convenient and accurate evaluation of the presence or absence of a target nucleic acid or the expression level of a target gene in a biological sample, by nucleic acid amplification means employing an enzyme. The invention provides a method for preparation of a sample for use in nucleic acid amplification, to be used for amplification of nucleic acid in a biological sample, which method comprises an extraction step in which a nucleic acid extraction reagent is added to the biological sample to obtain a nucleic acid extract, and a purification step in which the nucleic acid extract is contacted with zeolite to obtain a nucleic acid amplification sample suitable for nucleic acid amplification, to allow nucleic acid detection at high sensitivity.