COVID-19 RT-LAMP Primer Mixture for Faster Accurate Diagnosis
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Solution Overview
Problem
Existing COVID-19 diagnosis methods, such as RT-PCR, require complex laboratory setups and long processing times, while isothermal amplification methods like RT-LAMP suffer from false positive results and lack of widespread adoption due to insufficient portability and technical simplicity.
Innovation Solution
A primer mixture comprising specific primers (SEQ ID Nos: 1-6) and additives (DMSO, Tween 20) for isothermal amplification, enhancing specificity and accuracy, enabling rapid and reliable COVID-19 detection using a simple heater and visual or fluorescence evaluation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If RT-PCR method is used for COVID-19 diagnosis, then diagnostic accuracy is maintained, but processing time increases to 60-90 minutes and device complexity increases
Solution Approach 1:
The patent changes the thermal cycling parameters to isothermal conditions at 65°C, eliminating the need for complex thermal cycling while maintaining diagnostic accuracy. This parameter change reduces processing time to 30 minutes while preserving measurement precision through optimized primer and probe sequences specific to SARS-CoV-2.
Solution Approach 2:
The patent replaces the mechanical thermal cycling system with a simple heating block or water bath operating at constant temperature. This substitution eliminates complex thermal cycling machinery while maintaining diagnostic accuracy through isothermal amplification conditions optimized for SARS-CoV-2 detection.
2Reliability
If RT-PCR method is used for COVID-19 diagnosis, then diagnostic reliability is maintained, but device complexity and infrastructure requirements increase
Solution Approach 1:
The patent replaces complex RT-PCR thermal cyclers with simple isothermal heating devices such as heating blocks or water baths. This substitution maintains diagnostic reliability through optimized isothermal amplification conditions while eliminating the need for complex laboratory infrastructure and specialized equipment.
Solution Approach 2:
The patent employs disposable reaction mixtures and primers designed for single-use isothermal amplification. These disposable components maintain diagnostic reliability through pre-optimized sequences while eliminating the need for expensive, complex equipment and extensive laboratory infrastructure.
3Loss of time
If isothermal amplification method is used for COVID-19 diagnosis, then processing time is reduced to 30 minutes and device simplicity is improved, but false positive rate increases
Solution Approach 1:
The patent applies local quality by designing primers and probes with specific sequences targeted to unique regions of the SARS-CoV-2 genome. This localized specificity at the molecular level ensures accurate viral detection while maintaining the speed advantages of isothermal amplification, thereby reducing false positives.
Solution Approach 2:
The patent optimizes isothermal amplification parameters including temperature (65°C), primer concentrations, and reaction buffer conditions to enhance specificity. These parameter changes maintain the rapid 30-minute processing time while reducing false positive rates through carefully controlled isothermal conditions.
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 mixture reduces false positives, provides high specificity, and delivers results twice as fast as RT-PCR, facilitating widespread on-site COVID-19 diagnosis without complex equipment.
Implementation Method 1
The isothermal amplification method, also known as RT-LAMP, allows the detection of the Sars-CoV-2 virus that causes Covid-19 with a testkit and heater
Implementation Method 2
the presence of the virus can be easily observed with the eye or a plate reader that can measure between 440-560 nm at the end of the process via color change
Implementation Method 3
0.05%-0.2% tween 20
Implementation Method 4
0.5%-2% dimethylsulfoxide (DMSO)
Data Source
AI summary
The present invention relates to a reaction mixture suitable for use in the diagnosis of Covid-19 by isothermal amplification method.