Dermatophyte PCR Primer Pair for Rapid Species Identification
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Solution Overview
Problem
Current methods for diagnosing dermatophyte skin, hair, and nail infections are inadequate, particularly in distinguishing closely related strains from the genus Trichophyton and Microsporum, leading to inaccurate diagnoses and prolonged treatment times due to low sensitivity and time-consuming culture-based identification methods.
Innovation Solution
A primer pair and nucleic acid sequence specific to pathogens like Trichophyton species, capable of amplifying and hybridizing with metalloprotease-related sequences, are used for PCR-based diagnosis, allowing for rapid and specific identification of dermatophytes on microarrays with fluorescent labeling for detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Speed
If direct microscopic examination is used for diagnosis, then the method is rapid and cheap, but sensitivity is low and false negative results occur in up to 15% of cases
Solution Approach 1:
The patent replaces the mechanical/optical direct microscopic examination system with a PCR-based molecular detection system. This substitution enables specific DNA amplification of dermatophyte sequences, achieving both rapid results (within hours) and high diagnostic accuracy by detecting fungal genetic material directly from clinical samples without requiring culture or direct visualization.
Solution Approach 2:
The patent introduces PCR amplification as an intermediary step between sample collection and diagnosis. By using specific primers to amplify dermatophyte DNA sequences, the method creates a detectable intermediate product that enhances sensitivity while maintaining rapid turnaround, resolving the contradiction between speed and reliability.
2Measurement precision
If culture-based identification is used, then specific species identification is achieved in 95% of cases, but diagnosis time extends to 10-15 days or up to 3-6 weeks for slow-growing isolates
Solution Approach 1:
The patent performs preliminary DNA extraction and PCR amplification directly from clinical samples without waiting for fungal culture growth. By targeting conserved dermatophyte DNA sequences, the method achieves specific species identification within hours, eliminating the 10-15 day or longer waiting period associated with traditional culture-based identification while maintaining diagnostic accuracy.
3Reliability
If conventional PCR methods are used, then fungal detection is possible, but distinction of closely related strains from Trichophyton and Microsporum genera is not reliable
Solution Approach 1:
The patent applies local quality by designing species-specific primers that target unique DNA sequences in different dermatophyte species. The primer pairs are designed to amplify specific regions of the ITS1 and ITS2 spacers that contain species-specific variations, enabling reliable differentiation of closely related strains from Trichophyton and Microsporum genera while maintaining overall fungal detection capability.
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
This approach enables rapid, specific, and reliable differentiation of closely related dermatophyte species, improving diagnostic accuracy and reducing treatment delays by facilitating quicker identification of pathogens like Trichophyton tonsurans, T. equinum, T. interdigitale, T. benhamiae, and T. concentricum.
Implementation Method 1
A primer pair and nucleic acid sequence specific to pathogens like Trichophyton species, capable of amplifying and hybridizing with metalloprotease-related sequences, are used for PCR-based diagnosis
Implementation Method 2
allowing for rapid and specific identification of dermatophytes on microarrays with fluorescent labeling for detection
Implementation Method 3
allowing for rapid and specific identification of dermatophytes on microarrays with fluorescent labeling for detection
Data Source
Figure 1
Figure 2A~2B
Figure 2C~2D
AI summary
The present invention relates to a primer pair comprising a forward primer and a reverse primer for amplifying a nucleic acid from a pathogen associated with a skin, hair and nail infection comprising SEQ ID NO22, a nucleic acid capable of hybridizing specifically to a nucleic acid sequence from a pathogen associated with a skin, hair and nail infection comprising SEQ ID NO22, a carrier comprising the nucleic acid, a method comprising the step detecting in a sample a nucleic acid sequence comprising SEQ ID NO22 from a pathogen associated with a skin, hair and nail infection, a use of the primer pair, the nucleic acid or the carrier for the diagnosis of a disease and a kit comprising the primer pair, the nucleic acid and/or the carrier for the diagnosis of a disease.