LAMP Primer Set for Rapid Candida Auris Detection
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Solution Overview
Problem
Existing methods fail to accurately and rapidly detect Candida auris, a pathogenic fungus, due to its similarity with closely related species, posing challenges in diagnosis, treatment, and controlling hospital-associated infections.
Innovation Solution
A primer set comprising FIP, BIP, F3, and B3 primers, along with optional loop primers, is used in a LAMP method for specific amplification of Candida auris DNA, enabling rapid and accurate detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional PCR or sequencing methods are used for Candida auris detection, then detection accuracy can be achieved, but detection time is excessively long and complexity increases
Solution Approach 1:
The patent replaces conventional PCR thermal cycling mechanical system with the LAMP isothermal amplification system that operates at constant temperature (65°C), eliminating the need for complex thermal cycler equipment and significantly reducing detection time while maintaining high accuracy through specific primer design
Solution Approach 2:
The patent changes the detection parameter from temperature-cycling-based PCR to isothermal LAMP amplification at 65°C, and uses turbidity measurement instead of gel electrophoresis or sequencing, enabling rapid detection within 60 minutes while preserving detection accuracy
2Reliability
If conventional PCR methods are used, then detection capability is achieved, but device complexity and operational difficulty increase
Solution Approach 1:
The patent replaces complex thermal cycler equipment with a simple water bath or heating block that maintains constant 65°C temperature, and substitutes gel electrophoresis apparatus with simple turbidity measurement using a spectrophotometer or even visual observation, dramatically reducing device complexity while maintaining reliable detection
Solution Approach 2:
The LAMP reaction system automatically produces visible turbidity changes that indicate positive results without requiring additional analysis equipment or complex operational steps, enabling non-experts to perform and interpret检测结果 reliably
3Measurement precision
If species-specific detection methods are developed, then detection specificity improves, but applicability to multiple Candida species decreases
Solution Approach 1:
The patent designs a universal LAMP primer set targeting the ITS1 region that can detect multiple Candida species (C. auris, C. albicans, C. tropicalis, C. glabrata) simultaneously, making the detection system adaptable to various species while maintaining high specificity through careful primer design that distinguishes between species based on sequence variations
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 method allows for rapid and specific detection of Candida auris within 20-60 minutes, distinguishing it from related species with high sensitivity and accuracy, even in contaminated specimens.
Implementation Method 1
A primer set comprising FIP, BIP, F3, and B3 primers, along with optional loop primers, is used in a LAMP method for specific amplification of Candida auris DNA
Data Source
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AI summary
A method for detecting Candida auris, including subjecting a nucleic acid sample obtained from a specimen to a nucleic acid amplification reaction by the LAMP method and detecting an amplification product, wherein, in the primer set to be used in the LAMP method, FIP is a polynucleotide including 5 to 20 nucleotides located on the 5'-terminal side and 5 to 20 nucleotides located on the 3'-terminal side in the nucleotide sequence represented by SEQ ID NO: 1, BIP is a polynucleotide including 5 to 20 nucleotides located on the 5'-terminal side and 5 to 20 nucleotides located on the 3'-terminal side in the nucleotide sequence represented by SEQ ID NO: 2, F3 is a polynucleotide including SEQ ID NO: 3, and B3 is a polynucleotide including SEQ ID NO: 4.