Membrane Assay Nucleic Acid Cleavage
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Solution Overview
Problem
Rapid diagnostic assays, especially those using small pore size membranes, face challenges with membrane blockage and flow inhibition due to cellular nucleic acid, particularly DNA, leading to reduced accuracy and increased background signals, which hampers the balance between speed, sensitivity, and reliability.
Innovation Solution
The method involves treating cell-containing samples with cell lysis agents like detergents followed by nucleic acid cleavage using physical, chemical, or biological means to break down DNA, allowing for improved fluid flow and reduced background signals through membranes of small pore size, thereby enhancing assay performance.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If small pore size membranes are used for rapid diagnostic assays, then sensitivity and concentration capability are improved, but membrane blockage and flow inhibition occur due to cellular nucleic acid
Solution Approach 1:
The patent applies preliminary action by treating the sample with nucleic acid-cleaving conditions before the sample contacts the membrane. This pre-treatment breaks down DNA and RNA into smaller fragments that cannot block the membrane pores, preventing flow inhibition before it occurs while maintaining the sensitivity benefits of small pore membranes
Solution Approach 2:
The patent extracts the harmful nucleic acid component from the sample by using enzymes (nucleases) to specifically target and break down DNA and RNA. This removal of the problematic substance allows the membrane to function properly without blockage while preserving the analyte of interest for detection
2Productivity
If cell lysis is performed to allow sample passage through membrane, then cell-containing samples can be processed, but nucleic acid release causes membrane blockage
Solution Approach 1:
The patent establishes continuity of useful action by combining cell lysis and nucleic acid degradation in a continuous process. The sample undergoes lysis to release intracellular components, and simultaneously or subsequently, nucleases continuously degrade the released nucleic acids, ensuring uninterrupted flow through the membrane while maintaining productivity
Solution Approach 2:
The patent introduces nucleases as intermediary agents that mediate between the cell lysis process and membrane flow. These enzymes act as a bridge that converts the harmful effect of released nucleic acid into a beneficial outcome by degrading it into non-blocking fragments, allowing both lysis and membrane flow to coexist
3Speed
If detergent lysis is used to break down cell membranes, then sample flow is improved, but background signals increase and accuracy decreases
Solution Approach 1:
The patent applies parameter changes by modifying the chemical composition of the lysis buffer to include nucleic acid-degrading enzymes. This change in buffer parameters (adding nucleases, adjusting pH, ionic strength) enables both rapid cell lysis and simultaneous nucleic acid degradation, achieving fast assay speed while maintaining accuracy by reducing background signals
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 improves the flow through membranes, increases the sample volume processed, and enhances sensitivity and accuracy by reducing membrane blockage and background signals, allowing for faster and more reliable assays.
Implementation Method 1
the cell (e.g. blood cell) membranes are fragmented and typically solubilised, for example by using detergents
Implementation Method 2
subjecting the thus-generated lysed sample to conditions causing the cleavage of nucleic acid molecules, especially DNA molecules
Data Source
AI summary
Provided herein is technology relating to methods for assaying a cell-containing body sample and particularly, but not exclusively, to treating a sample under conditions to cause cell lysis, preferably by means of a detergent; and subjecting the lysed sample to conditions causing the cleavage of nucleic acid molecules. The technology additionally relates to using nucleic acid cleavage conditions to enhance a membrane assay, a device for carrying out such an assay, and a kit for use in the assay.