Reagent Composition for Direct Amplification from Crude Biological Samples
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Solution Overview
Problem
Current methods for detecting nucleic acids from microbes or infectious agents in crude biological samples are complex, time-consuming, and prone to errors due to the need for multiple steps, potential damage to nucleic acids during inactivation processes, and interference from sample components, leading to misdiagnoses and resource inefficiencies.
Innovation Solution
A treatment solution comprising a surfactant, protease, chelating agent, and buffering salt is used to inactivate and disrupt viral particles directly in crude samples, allowing direct nucleic acid detection without extraction, thereby reducing processing steps and improving detection efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If nucleic acid extraction is performed before detection, then detection accuracy is improved, but processing time and complexity increase
Solution Approach 1:
The patent combines the nucleic acid extraction step with the detection step by using a treatment solution that performs both functions in a single unified process, eliminating the need for separate extraction and detection procedures
Solution Approach 2:
The treatment solution is designed to perform multiple functions simultaneously: it inactivates viral particles, releases nucleic acids, and creates optimal conditions for detection, thereby serving as a multi-functional reagent that replaces multiple separate steps
2Reliability
If multiple processing steps are used, then detection reliability is improved, but device complexity and personnel demands increase
Solution Approach 1:
Multiple processing functions (inactivation, lysis, nucleic acid release, and detection optimization) are merged into a single treatment solution that can be applied in one step, reducing the number of separate processing steps while maintaining detection reliability
Solution Approach 2:
The treatment solution is formulated as a universal reagent that handles multiple aspects of sample preparation and detection optimization simultaneously, reducing the need for multiple specialized reagents and procedures
3Object-affected harmful factors
If viral inactivation is performed aggressively, then safety is improved, but nucleic acid integrity deteriorates
Solution Approach 1:
The treatment solution uses optimized chemical parameters (pH, temperature, reagent concentrations) that enable effective viral inactivation while maintaining conditions favorable for nucleic acid stability and integrity throughout the process
4Measurement precision
If sample components are removed, then detection precision is improved, but processing time and substance loss increase
Solution Approach 1:
The treatment solution acts as an intermediary that chemically modifies the interaction between sample components and the detection system, neutralizing interference from substances like hemoglobin and bilirubin without requiring their physical removal
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 enables faster, more reliable detection of nucleic acids with reduced personnel and infrastructure demands, providing stable and sensitive results even in crude samples, and allowing for flexible processing conditions.
Implementation Method 1
A treatment solution comprising a surfactant, protease, chelating agent, and buffering salt is used to inactivate and disrupt viral particles directly in crude samples
Implementation Method 2
A treatment solution comprising a surfactant, protease, chelating agent, and buffering salt is used to inactivate and disrupt viral particles
Implementation Method 3
A treatment solution comprising a surfactant, protease, chelating agent, and buffering salt is used to inactivate and disrupt viral particles
Data Source
AI summary
Disclosed are compositions, assays, methods, diagnostic methods, kits and diagnostic kits for the detection of target nucleic acids, including those from microbes and/or from infectious agents such as SARS-CoV-2 and other viruses. Embodiments described herein are designed to enable processing and analysis of the sample to detect target nucleic acids within the sample without requiring extraction and/or isolation of nucleic acid from the sample prior to subsequent processing steps. Samples analyzed can thus be “crude” biological samples that do not require pre-processing prior to placement in the workflow.