RAA Primer Probe Combination for Candida Auris Detection
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Solution Overview
Problem
Current diagnostic tools for detecting Candida auris are costly, have low accuracy, and often lead to misdiagnosis, making it difficult to timely prevent and control the spread of this highly infectious and drug-resistant fungus.
Innovation Solution
A constant temperature nucleic acid amplification RAA primer probe combination is developed, comprising specific upstream and downstream primers and a probe, which can accurately detect Candida auris without cross-reacting with other pathogenic bacteria, and can be used in a simple, cost-effective kit for rapid detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional phenotypic identification and microorganism identification systems are used, then the identification process is simple, but the accuracy is low leading to misdiagnosis
Solution Approach 1:
The patent extracts the specific identification function from complex commercial systems by designing primers and probes that specifically target Candida auris genetic sequences. This allows the detection function to be separated from the complex equipment and databases required by commercial systems, achieving high accuracy through simple nucleic acid amplification.
Solution Approach 2:
The patent uses disposable nucleic acid amplification reagents (primers and probes) that are inexpensive compared to commercial identification systems. These single-use reagents eliminate the need for expensive, maintainable equipment while providing accurate identification through specific genetic targeting.
2Measurement precision
If MALDI-TOF MS is used for identification, then the identification accuracy can be improved, but the cost and equipment requirements increase
Solution Approach 1:
The patent replaces the mechanical mass spectrometry system (MALDI-TOF MS) with a biochemical nucleic acid amplification system. This substitution maintains identification accuracy through specific genetic targeting while eliminating the need for expensive mass spectrometry equipment, making the detection accessible to resource-limited settings.
Solution Approach 2:
The patent uses inexpensive, disposable nucleic acid reagents instead of expensive MALDI-TOF MS equipment. The primers and probes can be synthesized at low cost and used in simple amplification reactions, providing the same identification capability without the high equipment investment and maintenance requirements.
3Measurement precision
If commercial detection systems are used, then the identification capability is improved, but the operation complexity and professional training requirements increase
Solution Approach 1:
The patent extracts the core detection function from complex commercial systems by using simple nucleic acid amplification with specific primers and probes. This eliminates the need for complex software databases, protein spectral analysis, and professional interpretation, making the operation simple enough for non-experts while maintaining high accuracy through genetic specificity.
4Loss of time
If conventional culture methods are used, then the equipment requirements are low, but the detection time and labor requirements increase
Solution Approach 1:
The patent changes the detection parameter from phenotypic observation (culture morphology) to genotypic detection (nucleic acid sequences). This parameter change enables rapid amplification and detection of specific genetic markers, reducing detection time from days to hours while using simple equipment that does not require complex infrastructure.
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 RAA primer probe combination achieves high specificity and sensitivity, allowing for the detection of Candida auris genomic DNA as low as 101 fg/reaction, and can be used in resource-limited settings without the need for expensive equipment or trained professionals, providing a rapid and reliable method for on-site detection.
Implementation Method 1
the recombinase-ssDNA complex binds to the dsDNA template to invade and form a D-loop structure
Implementation Method 2
in the presence of a ssDNA-binding protein
Implementation Method 3
a DNA polymerase to extend the ssDNA
Implementation Method 4
The recombinase aided amplification (RAA) method refers to a nucleic acid constant temperature amplification technology that can achieve single-molecule nucleic acid detection at normal temperature within 20 minutes
Data Source
AI summary
The present invention discloses a constant temperature nucleic acid amplification RAA primer probe combination for detecting Candida auris and use thereof are provided. The primer probe combination has high amplification efficiency, strong specificity, and no obvious false positive, and can effectively distinguish Candida auris from three closely related bacteria frequently misdiagnosed in a clinical microorganism identification system. Based on an RAA-lateral flow chromatography technology, the present invention establishes a method for rapidly detecting Candida auris can be used for detecting an actual sample after passing the specificity and sensitivity evaluation, and provides a rapid, sensitive, reliable, and effective new method for on-site rapid detection of Candida auris. The method does not need complex instruments, and is particularly suitable for rapid screening and detection of Candida auris in basic laboratories and quarantine sites.