E Gene RT-PCR Detection to Reduce Coronavirus False Negatives
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing real-time fluorescence RT-PCR methods for detecting novel coronavirus (2019-nCoV) are prone to missed tests due to RNA virus mutations, necessitating a more precise and sensitive detection system.
Innovation Solution
A primer pair set and probe set targeting the 2019-nCoV genome envelope protein (E) gene, along with an internal standard system, are developed to enhance detection specificity and prevent false negatives.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional RT-PCR detection methods targeting ORF1ab and N genes are used, then detection can be performed with existing protocols, but missed tests occur due to RNA virus mutations
Solution Approach 1:
The patent divides the detection system into multiple independent primer pairs targeting different genes (ORF1ab, N, and E genes). This segmentation allows the system to detect mutations in different regions independently, preventing missed tests due to viral mutations while maintaining overall detection reliability.
Solution Approach 2:
The patent introduces a new detection parameter by adding detection of the E gene as a third target alongside the traditional ORF1ab and N genes. This parameter change expands the detection coverage to include additional viral genes, improving both reliability and precision by providing more targets for accurate detection.
2Reliability
If multiple primer pairs targeting different genes are used to prevent missed detections, then detection reliability improves, but system complexity increases
Solution Approach 1:
The patent merges multiple primer pairs targeting different genes (ORF1ab, N, and E) into a single RT-PCR detection system that processes all targets simultaneously in one reaction. This combining approach improves detection reliability by providing multiple targets while avoiding the complexity of separate detection systems, as all primers work together in a unified assay.
3Measurement precision
If targeted detection of the E gene is implemented, then false negatives from mutations are reduced, but detection cost increases
Solution Approach 1:
The patent designs the E gene primer pairs to serve multiple functions: detecting the E gene specifically, identifying mutations, and preventing false negatives. This multi-functionality allows a single detection system to achieve improved precision through E gene targeting while avoiding the need for entirely separate detection systems, thereby controlling costs.
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 system provides high-efficiency, low-cost detection of 2019-nCoV with improved accuracy by targeting the E gene, reducing false negatives and confirming missed detections caused by mutations.
Implementation Method 1
Real-time fluorescence RT-PCR is one of the common methods for coronavirus nucleic acid detection
Implementation Method 2
The method has high sensitivity and good specificity, which can accurately and quickly provide the test results of the target virus
Implementation Method 3
the 5′ end of each probe is labeled with a fluorescent reporter group; and/or, the 3′ end of each probe is labeled with a fluorescence quenching group
Data Source
AI summary
A real-time fluorescence RT-PCR detection method and a kit for a novel coronavirus 2019-nCoV. Specifically, the present invention relates to a kit and a method for detecting the nucleic acid of an E gene of the novel coronavirus 2019-nCoV. The kit and the method have extremely high sensitivity and specificity, and can significantly improve the accuracy of virus identification.