Multi-Gene SARS-CoV-2 Primer Sets for Sensitive PCR Detection
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Solution Overview
Problem
The timely detection of SARS-CoV-2 infections is crucial for effective therapeutic intervention, as the virus is highly contagious and can be asymptomatic, necessitating early detection to prevent spread and enable timely treatment.
Innovation Solution
The use of specific oligonucleotide primer pairs and probes for nucleic acid amplification and detection, including probe-free and probe-based PCR methods, to identify SARS-CoV-2 in samples, along with the development of biomarkers for drug targets and therapeutics.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional virus detection methods are used, then detection can be performed, but the detection speed and sensitivity are insufficient for timely therapeutic intervention
Solution Approach 1:
The patent segments the virus detection process into specific target identification (N gene, E gene, S gene, ORF1ab gene) using targeted primer pairs. This segmentation allows parallel detection of multiple viral components, improving both sensitivity and speed compared to conventional single-target methods
Solution Approach 2:
The patent optimizes detection parameters including primer annealing temperatures (50-65°C), extension temperatures (72°C), and cycle numbers (35-45 cycles) to enhance amplification efficiency. These parameter changes enable faster and more sensitive detection while maintaining specificity
2Reliability
If early detection methods are implemented, then timely treatment can be initiated, but the complexity of detection procedures increases
Solution Approach 1:
The patent provides a universal detection system with multiple primer pairs that can detect different SARS-CoV-2 genes (N, E, S, ORF1ab) using the same basic RT-PCR platform. This multi-functionality allows a single laboratory setup to perform comprehensive viral detection without requiring multiple specialized systems
Solution Approach 2:
The patent uses nucleic acid amplification to create copies of viral genetic material, making the undetectable visible. The RT-PCR process generates millions of copies of target sequences, enabling reliable detection of even trace viral amounts while maintaining procedural simplicity
3Measurement precision
If specific primer pairs are used for SARS-CoV-2 detection, then detection specificity is improved, but the ability to detect variant strains may be reduced
Solution Approach 1:
The patent targets multiple distinct viral genes (N, E, S, ORF1ab) with separate primer pairs, so that if one target shows variability, others can still be detected. This segmentation across multiple genomic regions maintains both specificity for SARS-CoV-2 and adaptability to variants
Solution Approach 2:
The primer pairs are designed to target highly conserved regions within each gene, making them universally applicable across different SARS-CoV-2 variants. The multi-gene approach ensures that at least some targets remain detectable even if mutations occur in specific regions
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
Enables sensitive and specific detection of SARS-CoV-2, facilitating early identification and isolation of infected individuals, supporting large-scale assays, and providing biomarkers for vaccine and drug development.
Implementation Method 1
preparing DNA from RNA by reverse transcriptase reaction
Implementation Method 2
performing nucleic acid amplification reaction employing an oligonucleotide primer pair to produce an amplicon
Implementation Method 3
employing an oligonucleotide primer pair selected from the group consisting of a nucleic acid sequence as set forth in SEQ ID NO: 6 and SEQ ID NO: 7
Implementation Method 4
detecting the presence or absence of the amplicon
Data Source
AI summary
A method of detecting the presence of SARS-CoV-2 using oligonucleotide primer pairs is disclosed. Oligonucleotide primer pairs for the detection of SARS-CoV-2 using polymerase chain reaction (PCR)-based methods are also disclosed. Further, biomarkers and probes for SARS-CoV-2 detection are disclosed. The disclosure also relates to the identification and isolation of biomarkers for use as probes for the detection of SARS-CoV-2 and further use of the biomarkers as a target for the development of drug targets and therapeutics for SARS-CoV-2.