Microbial Genomic Detection Using DIANAs for Fast Bloodstream Diagnosis
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Solution Overview
Problem
Current methods for identifying microbial pathogens in bloodstream infections are slow, insensitive, and lack broad coverage, making it difficult to achieve early and accurate disease management.
Innovation Solution
The use of DNA Invading Artificial Nucleic Acids (DIANAs) for rapid and specific detection of microbial genetic materials, allowing for the identification and evaluation of microorganisms in a sample without culturing, and providing detailed information on microbial species and their resistance to antimicrobials.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If traditional blood-culture methods are used to identify microbial pathogens, then the identification can be performed with simple equipment, but the detection time is prolonged (taking up to several days)
Solution Approach 1:
The patent replaces traditional mechanical/cultural detection methods with molecular genetics-based detection. Specifically, it uses PCR amplification and DNA hybridization to detect microbial pathogens directly from blood samples, eliminating the need for prolonged bacterial culture growth. This substitution of detection mechanism reduces identification time from days to hours while maintaining or improving accuracy through direct genetic material detection.
Solution Approach 2:
The patent changes the detection parameter from measuring bacterial growth (colony formation) to detecting specific genetic sequences. By targeting conserved genetic regions across multiple pathogen types and using fluorescently labeled probes, the system achieves rapid detection without requiring the physical growth of organisms, thus reducing time loss while maintaining diagnostic reliability.
2Productivity
If molecular approaches are used to detect microbial pathogens, then detection speed is improved, but sensitivity and accuracy are insufficient for low pathogen loads
Solution Approach 1:
The patent applies preliminary action through pre-amplification of microbial genetic material before final detection. The method includes a nucleic acid amplification step that enriches the target sequences from the complex blood sample matrix, thereby concentrating the signal and improving detection sensitivity. This preliminary amplification enables the subsequent hybridization step to detect even low pathogen loads (1-100 CFU/ml) with high accuracy.
Solution Approach 2:
The patent uses fluorescently labeled DNA probes as intermediaries between the amplified genetic material and the detection system. These probes bind specifically to conserved regions of microbial DNA, providing a measurable signal that indicates pathogen presence. The use of fluorescent intermediaries enhances detection sensitivity by converting molecular binding events into detectable optical signals, enabling accurate detection at low pathogen loads.
3Adaptability or versatility
If broad-spectrum pathogen detection is attempted, then coverage is improved, but diagnostic detail and species-level identification are compromised
Solution Approach 1:
The patent segments the detection system into multiple parallel detection channels, each targeting specific conserved genetic regions of different pathogen groups. By using a panel of fluorescently labeled probes that can be simultaneously applied to a single sample, the system achieves broad pathogen coverage while maintaining the ability to distinguish between different species and groups through specific binding patterns. This segmentation allows multiplex detection without sacrificing diagnostic detail.
Solution Approach 2:
The patent creates a universal detection platform that can identify multiple pathogen types simultaneously using a single assay system. The method employs conserved genetic targets and fluorescent probe technology that work across diverse pathogen groups (bacteria, fungi, viruses), enabling one test to provide detailed species-level identification for numerous pathogens. This multi-functionality achieves both broad coverage and high diagnostic precision.
4Ease of operation
If pathogen detection is performed in complex blood matrices, then real clinical samples can be analyzed, but detection accuracy is reduced due to interference
Solution Approach 1:
The patent extracts and removes interfering components from the blood sample matrix before detection. The method includes steps to lyse eukaryotic cells and remove host DNA, thereby eliminating background noise and interference from the complex blood matrix. This extraction of interfering substances allows the subsequent PCR and hybridization steps to proceed with high accuracy, as the detection is performed on purified microbial genetic material rather than in the presence of competing biological molecules.
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, sensitive, and specific identification of microbial pathogens in blood samples, including their resistance profiles, overcoming the limitations of traditional methods by providing detailed microbial spectra and reducing the time required for diagnosis.
Implementation Method 1
contacting the amplified microbial genetic materials with a plurality of DNA Invading Artificial Nucleic Acids (DIANAs), wherein the plurality of DIANAs comprise one or more sequences selected from the group consisting of SEQ ID NOs: 20-571; and detecting binding of one or more of the plurality of DIANAs to the microbial genetic material of its respective single species or group of microbes
Data Source
AI summary
The present disclosure generally relates to the field of microbial pathogen detection and identification utilizing genomic sequence recognition.