Lysis Protection Solution for Stable RNA Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current nucleic acid detection kits for COVID-19 have high false negative rates and low sensitivity due to issues with primer and probe design, RNA degradation, and inefficient sample processing, leading to prolonged detection times and limited capacity in clinical settings.
Innovation Solution
A lysis/protection solution and specific primer/probe set are developed to stabilize RNA, allowing for one-step reverse transcription and amplification, enhancing sensitivity and specificity, and enabling rapid detection without the need for RNA extraction or purification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional primer and probe sets are used for nucleic acid detection, then the detection process is simpler, but the sensitivity is low and false negative rate is high
Solution Approach 1:
The detection system is segmented into multiple functional components: protection sequences (AP-WHN-1 and AP-WHORF1ab-1) that bind to specific viral RNA regions, lysis/protection solution for RNA stabilization, and optimized primers/probes. This segmentation allows each component to perform its specific function optimally, achieving 100 times higher sensitivity while maintaining procedural simplicity through integrated reagent formulation.
Solution Approach 2:
The patent employs composite reagent formulations combining lysis buffers, protection sequences, primers, and probes into integrated kits. The lysis/protection solution contains multiple chemical components working synergistically to stabilize RNA while the protection sequences specifically bind to viral RNA, creating a composite system that achieves both high sensitivity and operational simplicity.
2Measurement precision
If RNA extraction and purification steps are included, then detection accuracy improves, but detection time increases and operational complexity increases
Solution Approach 1:
The patent merges the lysis, protection, and detection preparation steps into a single integrated lysis/protection solution. This solution simultaneously stabilizes RNA and prepares it for direct amplification without requiring separate extraction and purification steps. The protection sequences bind to viral RNA in the lysate, protecting it from degradation while maintaining accessibility for primer binding, thereby achieving both accuracy and speed.
Solution Approach 2:
The lysis/protection solution performs preliminary RNA stabilization and protection actions before the amplification step. By pre-binding protection sequences to viral RNA during the lysis step, the system ensures RNA integrity is maintained throughout the subsequent detection process, eliminating the need for time-consuming purification steps while preserving detection accuracy.
3Reliability
If multiple target gene loci are detected to form compounds, then RNA stability improves and detection reliability increases, but the complexity of the detection system increases
Solution Approach 1:
The protection sequences AP-WHN-1 and AP-WHORF1ab-1 are designed to bind to multiple target gene loci of the novel coronavirus simultaneously. This multi-functionality allows a single reagent system to detect and protect multiple viral RNA targets, improving detection reliability through compound formation at multiple sites while avoiding the need for separate detection systems for each target.
Solution Approach 2:
The patent optimizes the concentration parameters of protection sequences (20 nM each) and adjusts the chemical composition of the lysis/protection solution to enable simultaneous binding to multiple target loci. By carefully controlling these parameters, the system achieves reliable multi-target detection and RNA stabilization without excessive complexity in the detection protocol.
4Measurement precision
If amplification cycles are increased to improve sensitivity, then detection sensitivity improves, but detection time increases
Solution Approach 1:
The patent achieves 100 times higher sensitivity through optimized parameter settings including protection sequence concentration (20 nM), primer concentrations, and amplification cycle conditions. The protected RNA compounds formed by AP-WHN-1 and AP-WHORF1ab-1 binding enable more efficient amplification, allowing high sensitivity to be achieved in fewer cycles compared to conventional methods, thereby reducing total detection time while maintaining or improving sensitivity.
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 solution achieves 100 times higher sensitivity and specificity, reducing detection time to 60-70 minutes, and allows for dual inactivation of the virus, improving detection capacity and safety in clinical settings.
Implementation Method 1
a plurality of target gene loci in the lysis/protection solution is effectively identified to form a compound, so that the 2019-nCoV RNA is more stable
Implementation Method 2
One-step reverse transcription is carried out on the enriched 2019-nCoV RNA compound to obtain cDNA
Implementation Method 3
the product is recognized by a self-designed 2019-nCoV specific probe, with 40 cycles signals thereof being amplified by a factor of 3.5×10^12
Data Source
AI summary
The present invention uses a lysis protection solution to treat patient samples, and the samples release genes targeting the 2019-nCoV virus. In the lysis protection solution, multiple target genetic loci are effectively identified by a protection sequence to form a compound, making 2019-nCoV RNA more stable and avoiding extracting purified RNA. The enriched 2019-nCoV RNA compound is further subjected to reverse transcription to obtain cDNA. A signal is then amplified for 40 cycles after the product is identified by a specific 2019-nCoV probe.