RNA Separation by Aqueous Partitioning for Intact Virus Detection
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Solution Overview
Problem
Existing methods struggle to effectively distinguish between intact virus particles capable of infection and free nucleic acids released from damaged or dead viruses, which is crucial for determining infectiousness in samples.
Innovation Solution
Utilizing an aqueous multi-phase partitioning system to separate free nucleic acids, such as RNA, preferentially into one phase while intact viruses distribute between or remain in another phase, allowing for the determination of infectious samples by analyzing specific phases.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional detection methods are used to identify viruses in biological samples, then virus detection can be performed, but free nucleic acids from damaged viruses cannot be distinguished from intact infectious viruses
Solution Approach 1:
The biological sample is segmented into distinct phases through aqueous two-phase partitioning, where intact viruses and free nucleic acids are separated into different phases based on their physical and chemical properties. This segmentation enables selective analysis of intact viruses without interference from free nucleic acids, resolving the detection precision issue.
Solution Approach 2:
Aqueous two-phase partitioning systems act as an intermediary mechanism that facilitates the separation of intact viruses from free nucleic acids. The partitioning system creates an intermediate state where different viral forms distribute differently between phases, enabling subsequent selective detection and solving the discrimination problem.
2Reliability
If samples are analyzed without separation to determine infectiousness, then analysis is rapid and simple, but accuracy in determining infectious capability is compromised
Solution Approach 1:
Aqueous two-phase partitioning is performed as a preliminary action before virus detection and analysis. This pre-separation step quickly distributes intact viruses and free nucleic acids into different phases, enabling subsequent focused analysis on the intact virus-containing phase. This preliminary separation improves reliability while minimizing time loss through efficient phase distribution.
Solution Approach 2:
The partitioning process exploits parameter changes in the sample environment to achieve separation. By adjusting pH, ionic strength, and polymer concentrations, the system optimizes the distribution of intact viruses versus free nucleic acids between phases, enhancing detection reliability without requiring excessive analysis time.
3Measurement precision
If free nucleic acids are not removed from samples, then sample processing is simpler and faster, but detection results are contaminated and less accurate
Solution Approach 1:
Free nucleic acids are extracted and removed from the sample matrix through aqueous two-phase partitioning, where they preferentially partition into one phase while intact viruses remain in another phase. This extraction eliminates contamination of detection results while maintaining relatively simple processing procedures, thus improving measurement precision without excessive complexity.
Solution Approach 2:
The system utilizes phase transitions in aqueous two-phase partitioning to separate free nucleic acids from intact viruses. By controlling the phase behavior of the partitioning system, free nucleic acids are directed into a specific phase that can be separately analyzed or discarded, improving detection accuracy while keeping the overall process manageable.
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 method enables the accurate differentiation between samples capable of infection and those that are not, providing a rapid and efficient means to identify potentially infectious individuals or specimens.
Implementation Method 1
partitioning a biological fluid comprising a virus and free nucleic acid in an aqueous multi-phase partitioning system
Data Source
AI summary
Certain aspects of the present disclosure generally relate to systems and methods for determining viruses such as coronaviruses. For instance, some aspects are directed to systems and methods for determining viruses using a partitioning system. Within the partitioning system, free RNA or other nucleic acids may preferentially partition into one phase, while intact viruses may be present in the other phase or in both phases. Accordingly, in some cases, free RNA or other nucleic acids may be preferentially removed, e.g., as compared to intact RNA or other nucleic acids present within a virus. In some cases, the phase containing intact viruses can be determined to determine the infectiousness, e.g., of a sample arising from a subject. This may be useful, for example, for distinguishing subjects who are capable of spreading an infection from those who are not infectious.