LAMP Primer Set for Rapid Trichophyton Gene Detection
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Solution Overview
Problem
Current methods for diagnosing tinea, particularly those involving direct microscopic examination and fungal culture, are cumbersome, require skilled techniques, and are slow, while PCR-based methods need specialized equipment and are not suitable for clinical settings.
Innovation Solution
A primer set comprising four specific oligonucleotides, including loop primers, designed for the LAMP method to amplify Trichophyton genes, allowing detection through turbidity or fluorescence measurement without requiring complex equipment or skilled labor.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If direct microscopic method or culture method is used for diagnosing tinea, then identification of fungus can be achieved, but the process is slow and requires skilled techniques
Solution Approach 1:
The patent replaces the mechanical/optical microscopic examination system with a molecular biology-based LAMP amplification system. By targeting specific ribosomal RNA genes of Trichophyton, the method achieves species-level identification through DNA amplification and detection, eliminating the need for skilled microscopic examination while significantly reducing diagnosis time from weeks to hours
Solution Approach 2:
The patent uses nucleic acid amplification to create multiple copies of the target Trichophyton gene sequences. The LAMP method amplifies specific ribosomal RNA gene regions, generating sufficient copies for detection without requiring culture growth, thus achieving rapid identification while maintaining high accuracy
2Measurement precision
If nested PCR or real-time PCR method is used for detecting Trichophyton, then detection sensitivity is improved, but specialized equipment and complex operations are required
Solution Approach 1:
The patent replaces the complex PCR thermal cycling system with an isothermal LAMP amplification system. By using temperature-independent DNA polymerase and carefully designed primer sets, the method achieves high detection sensitivity without requiring thermal cyclers, making it suitable for simple water bath or heat block incubation at constant temperature
Solution Approach 2:
The patent changes the temperature parameter from variable (PCR) to constant (LAMP). The LAMP method maintains amplification efficiency at a fixed temperature (typically 60-65°C), eliminating the need for complex temperature cycling equipment while achieving comparable or superior detection sensitivity through isothermal amplification kinetics
3Measurement precision
If fungal culture method is used, then fungal species identification can be achieved, but several weeks are required and skilled techniques are needed
Solution Approach 1:
The patent performs preliminary amplification of the target gene sequences before detection. By using LAMP primers that specifically target Trichophyton ribosomal RNA genes, the method pre-amplifies sufficient target material during a short incubation period, enabling rapid species identification without waiting for fungal culture growth over several weeks
Solution Approach 2:
The patent replaces the biological culture system with a molecular amplification system. Instead of relying on fungal growth and colony formation, the method directly amplifies and detects Trichophyton-specific DNA sequences, achieving species identification in hours rather than weeks and eliminating the need for culture maintenance skills
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 rapid and sensitive detection of Trichophyton species like T. rubrum and T. mentagrophytes in about two days, eliminating the need for skilled techniques and specialized equipment, and providing results comparable to conventional methods.
Implementation Method 1
the LAMP method is a method of amplifying DNA quickly at a fixed temperature
Implementation Method 2
allows the amplified DNA to be confirmed even by a visual check through measurement such as turbidity
Data Source
Figure 1
AI summary
The present invention has an object to provide a simpler and quicker method for diagnosing tinea, method for detecting a Trichophyton, or method for detecting the Trichophyton gene. Use of at least four kinds of specific primers each designed based on a DNA sequence of the Trichophyton gene makes it possible to simply and quickly detect the Trichophyton gene.