Dermatophyte Diagnosis Using Specific Primer Pairs
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Solution Overview
Problem
Current methods for diagnosing dermatophyte skin, hair, and nail infections are inadequate for rapid and specific distinction of closely related strains from the genus Trichophyton and Microsporum, leading to false negatives and prolonged diagnosis times.
Innovation Solution
A primer pair and nucleic acid probe capable of amplifying and specifically hybridizing to sequences from pathogens like Trichophyton species, allowing for rapid identification of strains such as T. tonsurans, T. equinum, T. interdigitale, T. benhamiae, and T. concentricum using PCR and microarray techniques.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
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/morphological examination system with a molecular biology system (PCR amplification and sequencing). Instead of visually examining spores and hyphae under a microscope, the invention uses nucleic acid amplification and sequence analysis to detect and identify dermatophytes, thereby maintaining rapid diagnosis while significantly improving sensitivity and accuracy
Solution Approach 2:
The patent creates molecular copies (amplified DNA sequences) of the target organism for analysis. By amplifying specific nucleic acid sequences from the dermatophyte sample and comparing them to reference sequences, the method enables highly sensitive detection without requiring direct observation of the organism, thus improving diagnostic reliability while maintaining speed
2Measurement precision
If culture method is used for specific species identification, then specificity is high, but diagnosis time is prolonged to 10-15 days or up to 3-6 weeks for slow growing isolates
Solution Approach 1:
The patent replaces the slow cultural growth system with a molecular amplification system. Instead of waiting for fungi to grow and develop characteristic morphology over days or weeks, the invention directly amplifies and sequences DNA from clinical samples, achieving species-level identification in hours while maintaining high accuracy
Solution Approach 2:
The patent performs preliminary molecular amplification and sequencing on clinical samples before culture is even attempted. By obtaining DNA sequences directly from the clinical specimen and comparing them to reference databases, the method provides rapid species identification without requiring the time-consuming growth phase that characterizes traditional culture methods
3Productivity
If PCR-based methods are used for fungal detection, then detection speed improves, but the ability to distinguish closely related strains from Trichophyton and Microsporum genera is insufficient
Solution Approach 1:
The patent applies local quality by selecting and analyzing specific regions of the fungal genome that contain strain-discriminatory information. Instead of using generic fungal detection primers, the invention targets variable regions within the ITS1 and ITS2 spacers that contain species- and strain-specific sequence variations, enabling precise differentiation of closely related dermatophytes while maintaining rapid PCR-based detection
Solution Approach 2:
The patent changes the analytical parameter from generic fungal detection to specific sequence variation analysis. By comparing nucleic acid sequences at the molecular level rather than relying on phenotypic characteristics or broad PCR detection, the method achieves both rapid detection and precise strain differentiation through sequence homology comparison with reference databases
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 specific diagnosis of dermatophyte infections by distinguishing between closely related strains, reducing false negatives and shortening diagnosis time through specific amplification and hybridization of nucleic acid sequences.
Implementation Method 1
A primer pair and nucleic acid probe capable of amplifying and specifically hybridizing to sequences from pathogens like Trichophyton species, allowing for rapid identification of strains
Implementation Method 2
A primer pair and nucleic acid probe capable of amplifying and specifically hybridizing to sequences from pathogens like Trichophyton species
Data Source
AI summary
A primer pair that includes a forward primer and a reverse primer is used to amplify a nucleic acid from a pathogen associated with a skin, hair and nail infection that includes SEQ ID NO: 22. A nucleic acid capable of hybridizing specifically to a nucleic acid sequence from a pathogen associated with a skin, hair and nail infection that includes SEQ ID NO: 22 is provided. A carrier that includes the nucleic acid is provided. A method can be used to detect in a sample a nucleic acid sequence including SEQ ID NO: 22 from a pathogen associated with a skin, hair, and nail infection. The primer pair, the nucleic acid, or the carrier may be useful for the diagnosis of a disease. A kit including the primer pair, the nucleic acid, and/or the carrier may be useful for the diagnosis of a disease.