Extracellular Nucleic Acid Stabilization in Blood Samples
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Solution Overview
Problem
Current methods for stabilizing extracellular nucleic acids in cell-containing biological samples, such as whole blood, are inefficient, leading to contamination with genomic DNA and degradation, particularly during storage and handling, which complicates diagnostic and prognostic applications.
Innovation Solution
A method involving the use of apoptosis inhibitors, hypertonic agents, and specific compounds to stabilize extracellular nucleic acids, preventing contamination and degradation, allowing for room temperature storage of whole blood samples for extended periods without compromising the integrity of the extracellular nucleic acid population.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional methods are used to separate cell-free fraction from cell-containing samples, then extracellular nucleic acids can be isolated, but cell breakage occurs during centrifugation which contaminates the sample with intracellular nucleic acids
Solution Approach 1:
The patent applies preliminary action by adding stabilizing agents (apoptosis inhibitors, hypertonic agents, and specific compounds) to the cell-containing sample before any separation process. This pre-stabilization prevents cell breakage and nucleic acid degradation during subsequent handling and storage, thereby preventing contamination with intracellular nucleic acids and maintaining the purity of extracellular nucleic acid isolation
2Ease of operation
If samples are stored at room temperature for extended periods, then ease of handling and shipping is improved, but extracellular nucleic acids degrade and become contaminated
Solution Approach 1:
The patent applies preliminary action by incorporating stabilizing agents into the sample collection tube before sample collection. These agents (apoptosis inhibitors, hypertonic agents, and specific compounds) are pre-positioned to immediately stabilize cells and extracellular nucleic acids upon sample collection, enabling reliable room temperature storage for extended periods while maintaining nucleic acid integrity
Solution Approach 2:
The patent uses stabilizing agents as intermediaries between the sample and the storage conditions. These agents mediate the interaction between extracellular nucleic acids and the storage environment, protecting the nucleic acids from degradation and contamination during room temperature storage and handling
3Measurement precision
If multiple centrifugation steps are performed to remove all cells, then contamination is reduced, but the process becomes tedious and time-consuming
Solution Approach 1:
The patent applies preliminary action by stabilizing cells and preventing cell breakage before any centrifugation steps. This pre-stabilization ensures that even if residual cells are present after a single centrifugation step, they remain intact and do not release intracellular nucleic acids, thereby achieving high purity isolation with minimal processing time
4Ease of operation
If EDTA is used as anticoagulant, then blood clotting is prevented, but it does not efficiently prevent cell degradation and nucleic acid contamination
Solution Approach 1:
The patent merges multiple functions into a single stabilizing composition that includes apoptosis inhibitors, hypertonic agents, and specific compounds in addition to or替代 EDTA. This combined formulation not only prevents blood clotting but also efficiently stabilizes cells, prevents cell degradation, and maintains extracellular nucleic acid integrity, thereby resolving the limitations of EDTA alone
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 stabilization method effectively reduces contamination with intracellular nucleic acids, maintains the integrity of extracellular nucleic acids, and allows for reliable diagnostic and prognostic analysis, even after prolonged storage at room temperature, enhancing the accuracy and reliability of nucleic acid testing.
Implementation Method 1
contacting the sample with a) at least one apoptosis inhibitor
Implementation Method 2
b) at least one hypertonic agent, which stabilizes the cells comprised in the sample
Implementation Method 3
The presence of elevated levels of extracellular nucleic acids such as DNA and/or RNA in many medical conditions... are protected from nucleases
Data Source
AI summary
The present invention provides methods, compositions and devices for stabilizing the extracellular nucleic acid population in a cell-containing biological sample using an apoptosis inhibitor, preferably a caspase inhibitor, a hypertonic agent and/or a compound according to formula (1) as defined in the claims.