Panfungal DNA Assays for Rapid Multiplex Detection
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Solution Overview
Problem
Current methods for detecting fungal infections are unreliable, time-consuming, and often provide false negative results, leading to delayed treatment and increased morbidity and mortality, especially in immunocompromised patients, and there is a lack of rapid tests for fungal presence in non-biological samples.
Innovation Solution
The use of DNA analysis for panfungal detection, including novel regions of fungal DNA and specific probes and primers for rapid identification of fungi, such as Candida, Aspergillus, and Pneumocystis, allowing for simultaneous detection of multiple species in a multiplex reaction.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If blood culture methods are used for fungal detection, then the detection can be performed with standard equipment, but the detection time is prolonged to 1-2 days and false negative results occur
Solution Approach 1:
The patent replaces the mechanical/biological culture system with a molecular biology system (PCR amplification and hybridization detection). Instead of relying on fungal growth in culture media over 1-2 days, the invention uses DNA extraction followed by specific probe hybridization to detect fungal presence directly, eliminating the time-consuming culture step while improving reliability through species-specific molecular targets
Solution Approach 2:
The patent performs preliminary DNA extraction and concentration of fungal genetic material before the actual detection step. By preparing the sample in advance and concentrating the target DNA, the method enables rapid subsequent detection without requiring prolonged incubation periods, thus reducing overall detection time while maintaining high sensitivity
2Reliability
If species identification is performed to determine fungal susceptibility to anti-fungal agents, then treatment can be optimized, but the identification process is time-consuming and delays treatment
Solution Approach 1:
The patent employs a universal probe that can detect and identify multiple fungal species simultaneously using a single assay system. This multi-functional approach allows clinicians to determine susceptibility to anti-fungal agents for various species (including Candida, Aspergillus, and others) without performing separate identification tests for each species, thereby optimizing treatment while minimizing time loss
3Productivity
If rapid detection tests are developed for fungal presence, then treatment can be initiated promptly, but the tests are currently unavailable for non-biological samples
Solution Approach 1:
The patent develops a universal detection system using conserved fungal DNA sequences that can detect fungi across diverse sample types. The methodology is adapted to work with both biological samples (blood, tissue) and non-biological samples (surfaces, equipment, environmental samples) by modifying the DNA extraction protocol while maintaining the same PCR and hybridization detection steps, thus enabling rapid treatment initiation without limiting sample type versatility
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 reliable detection of fungi in both biological and non-biological samples, facilitating timely treatment and ensuring accurate identification of fungal species, thereby reducing morbidity and mortality.
Implementation Method 1
The probe is capable of hybridizing to a target sequence
Implementation Method 2
The molecular beacon comprises a sequence capable of hybridizing to a target sequence and a detectable label
Data Source
AI summary
Methods and kits are described for testing for the presence or absence of any fungus in a sample. Examples of fungi that can be detected include, but are not limited to, those belonging to the genera Candida, Aspergillus and Pneumocystis. The methods include obtaining a sample suspected of containing fungal nucleic acid, including at least one universal region of fungal nucleic acid, and testing for the presence or absence in the sample of the at least one universal region of fungal nucleic acid. Samples may be biological or non-biological.