Virus Infectivity Detection Kit Using Photoactivatable Dye and Nuclease
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Solution Overview
Problem
Current methods for detecting virus infectivity, such as PCR/RT-PCR, cannot distinguish between live and dead viruses, leading to false positives and unnecessary quarantine of recovered patients, and existing antigen and antibody tests have low sensitivity and specificity.
Innovation Solution
A sample pretreatment kit and method using a photoactivatable dye and nuclease to differentiate between live and dead viruses by inhibiting nucleic acid amplification from dead cells, allowing for accurate determination of virus infectivity through PCR amplification differences.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If PCR/RT-PCR is used for virus detection, then detection sensitivity is improved, but the ability to distinguish infectious from non-infectious viruses deteriorates
Solution Approach 1:
The detection system is segmented into two independent reaction systems: one for detecting total viral nucleic acid (including both infectious and non-infectious viruses) and another for detecting only infectious viruses. This segmentation allows simultaneous acquisition of both sensitivity and infectivity information without compromising either detection capability.
Solution Approach 2:
A cell membrane integrity marker (such as PMA or EMA) is introduced as an intermediary substance that selectively penetrates dead cells but not live cells. This marker serves as a mediator to differentiate between infectious and non-infectious viruses during the PCR amplification process, enabling the system to preserve infectivity information while maintaining high detection sensitivity.
2Ease of manufacture
If antibody detection is used for virus detection, then detection cost is reduced and operation is simplified, but detection sensitivity and specificity deteriorate
Solution Approach 1:
The method performs preliminary differentiation of virus viability status before the main detection process by using cell membrane integrity markers. This preliminary action allows the subsequent antibody or antigen detection to focus specifically on infectious viruses, thereby improving both sensitivity and specificity while maintaining the cost-effectiveness and operational simplicity of serological methods.
3Productivity
If antigen detection is used for virus detection, then large-scale screening is facilitated, but false negative rate increases
Solution Approach 1:
The detection approach is segmented to separately identify infectious viruses from total viruses. By using cell membrane integrity markers in combination with antigen detection methods, the system can perform large-scale screening while reducing false negatives, as the marker-based differentiation adds specificity without significantly increasing operational complexity.
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 method effectively distinguishes infectious from non-infectious viruses, reducing false positives and improving diagnostic accuracy, enabling more precise patient management and reducing social burden.
Implementation Method 1
a photoactivatable dye capable of intercalating into a nucleic acid... exposing the test sample to a light for photoactivation
Implementation Method 2
a photoactivatable dye capable of intercalating into a nucleic acid
Implementation Method 3
a nuclease capable of degrading the nucleic acid
Data Source
AI summary
A sample pretreatment kit for detecting virus infectivity includes a photoactivatable dye capable of intercalating into a nucleic acid, and a nuclease capable of degrading the nucleic acid. The photoactivatable dye includes PMA dye, PMAxx dye, EMA, platinum compounds, or palladium compounds. The method for detecting virus infectivity includes steps of: (a) dividing a clinical sample into a test sample and a control sample; (b) treating the test sample with a photoactivatable dye and a nuclease; (c) exposing the test sample to a light for photoactivation; (d) amplifying a target nucleic acid in the test sample and the control sample; and (e) determining virus infectivity based on amplification results of the test sample and the control sample.