Nucleic Acid Stabilization Reagents for Direct PCR Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
The shortage of RNA extraction reagents during viral pandemics, such as the SARS-CoV-2 outbreak, hampers molecular diagnostics due to supply constraints and the labor-intensive, time-consuming nature of nucleic acid purification, which can lead to sample degradation and reduced sensitivity in detection methods like PCR and sequencing.
Innovation Solution
A method and reagents that allow for direct detection of nucleic acids from stabilized samples without the need for purification, using a transport solution containing chaotropic agents and chelating agents, and an activating solution with a buffered-based pH, enabling PCR amplification and detection of pathogens like SARS-CoV-2 without prior nucleic acid extraction.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If nucleic acid purification step is performed, then detection sensitivity is improved, but processing time and labor are increased
Solution Approach 1:
The invention extracts and removes the nucleic acid purification step from the traditional diagnostic workflow. By using stabilization reagents that contain chaotropic agents and chelating agents, the method eliminates the need for separate purification operations while maintaining detection sensitivity, thus reducing processing time and labor requirements.
Solution Approach 2:
The stabilization reagents are applied preliminarily at the sample collection stage to preserve nucleic acids and inhibit degradation. This preliminary action ensures that nucleic acids remain intact and detectable without requiring subsequent purification steps, thereby reducing overall processing time while maintaining sensitivity.
2Reliability
If nucleic acid purification step is performed, then sample quality is improved, but reagent consumption and cost are increased
Solution Approach 1:
The stabilization reagents serve multiple functions simultaneously: they stabilize nucleic acids, inhibit nucleases, prevent sample degradation, and enable direct PCR amplification. This multi-functionality eliminates the need for separate purification reagents and materials, reducing overall reagent consumption and cost while maintaining sample quality.
Solution Approach 2:
The invention merges the stabilization and detection preparation functions into a single reagent system. The stabilization reagents are designed to work directly with PCR master mix without requiring additional purification reagents, thereby reducing total reagent consumption while ensuring sample quality for reliable detection.
3Measurement precision
If nucleic acid purification step is performed, then detection accuracy is improved, but throughput and productivity are reduced
Solution Approach 1:
The method skips the time-consuming nucleic acid purification step by using stabilization reagents that preserve nucleic acids in a directly amplifiable state. This allows laboratories to rush through to the detection phase more quickly, significantly increasing throughput and productivity while maintaining detection accuracy through proper stabilization.
Solution Approach 2:
By performing nucleic acid stabilization preliminarily at sample collection, the method eliminates the need for subsequent purification steps. This preliminary action ensures detection accuracy is maintained while dramatically increasing laboratory throughput and productivity by reducing the number of processing steps required.
4Reliability
If nucleic acid purification step is performed, then sample integrity is improved, but manual handling and degradation risk are increased
Solution Approach 1:
The stabilization reagents provide self-service protection for nucleic acids by containing chaotropic agents and chelating agents that actively inhibit nuclease activity and prevent degradation. This self-protective mechanism reduces the need for careful manual handling during purification steps, minimizing degradation risk while maintaining sample integrity.
Solution Approach 2:
The invention extracts the manual purification steps from the workflow, eliminating the need for hands-on manipulation that can lead to sample degradation. By using stabilization reagents that preserve nucleic acids in place, the method reduces manual handling requirements while maintaining sample integrity through chemical protection.
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 enhances the efficiency and accuracy of molecular diagnostics by eliminating the purification step, allowing for faster processing and maintaining the structural integrity of nucleic acids, thereby improving the sensitivity and throughput of diagnostic tests.
Implementation Method 1
the transport solution comprises a chaotropic agent and a chelating agent, wherein the transport solution stabilizes a nucleic acid molecule of the pathogen at room temperature
Implementation Method 2
the transport solution comprises a chaotropic agent and a chelating agent, wherein the transport solution stabilizes a nucleic acid molecule of the pathogen at room temperature
Implementation Method 3
PCR amplification of the mixture to amplify the detectable polynucleotide
Data Source
AI summary
A method of use and reagent kit for nucleic acid stabilization and room temperature transport of SARS-CoV-2 and other respiratory viruses, followed by rapid identification are disclosed. Using the methods and compositions herein, molecular diagnostics or detection of SARS-CoV-2 and other respiratory viruses from the biological sample is performed without extraction or purification of viral DNA or RNA.