Multiplex Coronavirus Nucleic Acid Kit for Mutation-Resistant PCR Detection
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Solution Overview
Problem
Current nucleic acid detection kits for novel coronavirus 2019-nCOV lack multiplex detection capabilities, leading to variable performance and a high risk of false positives or negatives due to genetic mutations, and are not suitable for large-scale, efficient, and cost-effective detection.
Innovation Solution
A primer and probe combination for multiplex detection of novel coronavirus 2019-nCOV nucleic acid, including specific primer pairs and probes, along with an internal standard for quality control, to ensure accurate and simultaneous detection of multiple targets, reducing false positives and negatives.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If single-target detection reagents are used, then the detection process is simple, but the detection performance varies greatly and false positives/negatives occur due to lack of simultaneous multi-gene detection
Solution Approach 1:
The patent combines multiple primer pairs and probes targeting different genes (N gene, ORF1ab gene, E gene) into a single multiplex detection system. This merging allows simultaneous detection of multiple viral genes in one reaction, improving detection performance consistency and reducing false positives/negatives while maintaining operational simplicity through unified protocol
Solution Approach 2:
The detection kit is designed with universal applicability by incorporating multiple target genes into a single reagent system. The same kit can detect N gene, ORF1ab gene, and E gene simultaneously, making it a multi-functional tool that maintains simple operation while significantly improving detection reliability through cross-validation of multiple targets
2Reliability
If virus isolation and culture method is used, then high specificity and sensitivity are achieved, but the clinical detection time is too long and the process is cumbersome
Solution Approach 1:
The patent replaces the traditional virus isolation and culture method (which involves complex mechanical and temporal processes) with real-time fluorescence PCR technology. This substitution maintains high specificity and sensitivity through molecular targeting while dramatically reducing detection time from days to hours by directly detecting viral nucleic acids rather than cultivating the virus
3Measurement precision
If real-time fluorescence PCR technology is used, then higher sensitivity, specificity and precision are achieved, but the reagents lack simultaneous detection capability for multiple genes
Solution Approach 1:
The patent merges multiple detection targets (N gene, ORF1ab gene, E gene) into a single real-time fluorescence PCR reaction system with multiple primer pairs and probes. This combination enables simultaneous detection of multiple genes with high precision, maintaining the advantages of real-time fluorescence PCR while adding comprehensive multi-gene detection capability through coordinated amplification and fluorescent signaling
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 kit enables high-efficiency, specific, and cost-effective detection of novel coronavirus 2019-nCOV by simultaneously detecting multiple targets, verifying results through internal standards, and preventing false positives and negatives, providing a reliable basis for virus identification and control.
Implementation Method 1
the probe is digested and degraded by the exonuclease activity of the 5′-3′ end of Taq enzyme
Implementation Method 2
the fluorescent signal emitted by the reporter group is absorbed by the quencher group
Data Source
AI summary
Provided is a nucleic acid detection kit for novel coronavirus 2019-nCOV, and in particular, provided are a kit and a method for multiple detection of the nucleic acid of the novel coronavirus 2019-nCOV. Three nucleic acid targets of the novel coronavirus 2019-nCOV can be detected at the same time. False positives can be prevented by means of mutual authentication among different targets, and detection omissions, which may be caused by mutations, are confirmed, such that the accuracy of virus identification is significantly improved.