Stabilizing Cell-Free Nucleic Acids with Urotropin Buffer
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Solution Overview
Problem
Current methods for stabilizing cell-free nucleic acids and cells in biological samples, such as whole blood or urine, often require multiple ingredients and are not storage-stable, leading to issues like spontaneous lysis and degradation of nucleic acids during analysis.
Innovation Solution
A composition comprising a buffering compound, such as citrate buffer, and urotropin, which acts as an anticoagulant and chelating agent, is used to stabilize cell-free nucleic acids and cells by maintaining a pH of 3.5 to 7, preventing coagulation and lysis, and is storage-stable for up to eight months.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If multiple ingredients are used for stabilizing cell-free nucleic acids and cells, then the stabilization effect is improved, but the device complexity and manufacturing complexity increase
Solution Approach 1:
The patent combines multiple stabilizing agents (urotropin, EDTA, and buffer) into a single composition that provides both cell-free nucleic acid stabilization and cell lysis prevention. This merging of functions reduces the number of separate ingredients needed while maintaining effective stabilization.
Solution Approach 2:
The composition is designed to perform multiple functions simultaneously: urotropin stabilizes cell-free nucleic acids, EDTA prevents coagulation and stabilizes cells, and the buffer maintains pH. This multi-functional approach improves stabilization effectiveness without requiring separate additives for each function.
2Duration of action of stationary object
If storage time is extended for later analysis, then the duration of action is improved, but the stability of nucleic acids and cells deteriorates due to spontaneous lysis and degradation
Solution Approach 1:
The stabilizing composition is added to the blood sample immediately after collection, before any degradation or lysis can occur. This preliminary stabilization action preserves nucleic acid integrity and prevents cell lysis during subsequent storage, enabling extended storage times without loss of sample quality.
Solution Approach 2:
The composition provides protective cushioning against degradation mechanisms by incorporating antioxidants and stabilizing agents that prevent oxidative damage and nucleic acid degradation during storage, thereby maintaining sample integrity over extended periods.
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 composition effectively preserves the integrity and quantity of cell-free nucleic acids and cells, allowing for reliable analysis through hybridization, sequencing, or amplification without significant changes, even after extended storage, and prevents cell lysis, ensuring accurate results.
Implementation Method 1
at least one anticoagulant and/or at least one chelating agent, in particular at least one chelating agent for bivalent cations, preferably for calcium ions
Implementation Method 2
at least one buffering compound in aqueous solution that buffers to a pH value of 7 or below, preferably 3.5 to 7.0
Implementation Method 3
stabilizing cells contained within the sample, e.g. against a spontaneous lysis
Implementation Method 4
stabilizing cell-free nucleic acids contained in the biological sample, particularly for the further analysis
Data Source
AI summary
A method for stabilizing cell-free nucleic acids. The method includes providing a composition and applying the composition to a biological sample as a stabilizing agent for the cell-free nucleic acids contained in the biological sample. The composition includes at least one buffering compound that buffers to a pH value of 7 or below, at least one anticoagulant and urotropin in aqueous solution.