SARS-CoV-2 LAMP Primer Set for Rapid Isothermal Detection
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Solution Overview
Problem
Current methods for detecting SARS-CoV-2, such as RT-PCR, are time-consuming and lack promptness, necessitating the development of more efficient techniques for rapid nucleic acid amplification.
Innovation Solution
A primer set comprising LAMP primers targeting specific ORF regions in the SARS-CoV-2 genome, including Orf1b, OrfM, OrfN, and OrfS, for use in the RT-LAMP method, which allows for isothermal amplification and rapid detection of the virus.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If RT-PCR method is used for detecting SARS-CoV-2, then detection specificity is improved, but detection time increases (6-8 hours)
Solution Approach 1:
The patent changes the amplification method from RT-PCR (requiring thermal cycling) to LAMP (isothermal amplification at constant temperature). This parameter change in the reaction conditions enables rapid amplification without complex temperature cycling, reducing detection time to 7-10 minutes while maintaining specificity through carefully designed primers targeting SARS-CoV-2 specific regions (N, E, and ORF1ab genes)
Solution Approach 2:
The patent extracts and utilizes the key advantage of LAMP method (isothermal amplification capability) while eliminating its traditional drawback (lower specificity). By selecting highly specific SARS-CoV-2 target regions and designing optimized primers, the invention achieves both rapid isothermal amplification and high detection specificity, separating the speed benefit from the specificity compromise
2Speed
If LAMP method is used for rapid detection, then detection speed is improved, but risk of cross-reactivity with related viruses increases
Solution Approach 1:
The patent divides the detection into three separate LAMP reactions, each targeting a different SARS-CoV-2 specific region (N gene, E gene, and ORF1ab gene). This segmentation approach ensures that at least one target region will be specific to SARS-CoV-2 and not cross-react with related coronaviruses, while the multiplexed isothermal amplification maintains rapid detection speed
Solution Approach 2:
The patent designs a universal primer set that can detect multiple SARS-CoV-2 genomic regions simultaneously through isothermal amplification. The LAMP primers are designed to target conserved yet specific regions across different SARS-CoV-2 variants, providing broad coverage while maintaining specificity and preventing cross-reactivity with other coronaviruses
3Reliability
If conventional RT-PCR is used, then amplification reliability is improved, but operational complexity increases
Solution Approach 1:
The patent changes the thermal cycling parameters of conventional RT-PCR to isothermal conditions for LAMP amplification. This single parameter change simplifies the operational protocol by eliminating the need for complex temperature cycling programs, while the isothermal amplification maintains high reliability through optimized primer design and specific target region selection
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 primer set enables rapid and sensitive detection of SARS-CoV-2, reducing detection time to approximately 7-10 minutes with high specificity and sensitivity, comparable to conventional methods while avoiding cross-reactivity with related viruses.
Implementation Method 1
technologies using a LAMP method that uses a specific primer set is reported
Implementation Method 2
A primer set comprising LAMP primers targeting specific ORF regions in the SARS-CoV-2 genome, including Orf1b, OrfM, OrfN, and OrfS, for use in the RT-LAMP method, which allows for isothermal amplification and rapid detection of the virus
Data Source
AI summary
One embodiment according to the present invention relates to a primer set for detecting SARS-CoV-2, the set comprising a plurality of LAMP primers targeting one or more open reading frame (ORF) regions in a SARS-CoV-2 genome, wherein the one or more ORF regions are selected from the group consisting of Orf1b, OrfM, OrfN, and OrfS; and a technique relating to use of this primer set.