C. minuta DNA Detection for Complex Microbiota Samples
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Solution Overview
Problem
Current methods for detecting Christensenella minuta bacteria are time-consuming, require cultivable bacteria, and are not suitable for complex samples with multiple bacterial seeds, lacking sensitivity and specificity, especially in diagnosing associated diseases.
Innovation Solution
An in vitro method detecting a specific sequence between 1.921.147 bp and 2.014.152 bp of the C. minuta reference genome, utilizing primers and probes to identify and quantify C. minuta, enabling rapid and specific detection in various samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional culturing methods are used to detect C. minuta, then identification can be achieved, but the detection time is prolonged to 24-48 hours and requires pure bacterial culture
Solution Approach 1:
The patent replaces the mechanical culturing process with a molecular detection system using PCR amplification and sequencing. Instead of relying on bacterial growth (mechanical/biological process taking 24-48 hours), the invention uses biochemical amplification of specific DNA sequences to detect C. minuta presence rapidly, eliminating the time-consuming culturing step while maintaining identification accuracy.
Solution Approach 2:
The patent performs preliminary action by designing and using specific primers that target unique sequences in the C. minuta genome before detection. This pre-prepared specificity allows the detection system to immediately recognize and amplify C. minuta DNA upon exposure to the sample, without requiring prior culturing or preparation steps, thus reducing detection time while ensuring accurate identification.
2Measurement precision
If conventional culturing methods are used, then bacterial identification is possible, but the method requires cultivable bacteria and simple samples
Solution Approach 1:
The patent replaces the biological culturing system with a molecular detection system that works directly on extracted DNA. This substitution eliminates the requirement for bacteria to be cultivable and allows the method to handle complex samples containing multiple bacterial species, as the PCR primers specifically target C. minuta sequences regardless of other contaminants present in the sample.
Solution Approach 2:
The patent applies local quality by designing primers that target specific unique sequences within the C. minuta genome (such as the 16S rRNA gene regions). This localized specificity allows the detection method to identify C. minuta accurately even in complex samples with multiple bacterial seeds, as the primers bind only to the intended target sequences and not to other bacteria.
3Measurement precision
If molecular diagnosis with 16S DNA fragment is used, then sensitive and specific detection is achieved, but the cost is high and contamination risks increase
Solution Approach 1:
The patent applies local quality by targeting specific regions of the 16S rRNA gene that are unique to C. minuta rather than using universal 16S primers. This localized targeting maintains high sensitivity and specificity while reducing the risk of contamination from other bacteria, as the primers will not amplify non-target sequences. The approach simplifies the overall process by focusing amplification only on the relevant specific sequences.
4Adaptability or versatility
If universal 16S DNA primers are used, then broad bacterial detection is possible, but specificity for C. minuta is reduced due to similar sequences in other species
Solution Approach 1:
The patent resolves this contradiction by using local quality - designing primers that target specific conserved regions within the 16S rRNA gene that are unique to C. minuta. These primers bind to sequences that are present in all C. minuta strains but absent or different in other bacterial species, thereby maintaining species-specific specificity while still detecting the full range of C. minuta variants.
Solution Approach 2:
The patent applies segmentation by dividing the 16S rRNA gene into specific target regions and selecting the most discriminatory segments for primer design. By focusing on particular gene regions (such as V1-V9 hypervariable regions) that show the most sequence divergence between C. minuta and other species, the method achieves high specificity while maintaining the ability to detect various C. minuta strains.
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 provides rapid, sensitive, and specific detection of C. minuta, allowing for disease diagnosis and therapeutic regimen adaptation, and monitoring its evolution in the intestinal microbiota.
Implementation Method 1
utilizing primers and probes to identify and quantify C. minuta
Implementation Method 2
Molecular diagnosis also allows the identification of a bacterial seed and is based on the extraction and amplification of a specific gene fragment
Data Source
AI summary
The present invention relates to an in vitro method for detecting or diagnosing the presence of at least one bacterium of the species Christensenella minuta in a subject, through the detection of at least a portion of a sequence specific for the Christensenella minuta reference genome. The present invention also relates to methods and kits for identifying, and monitoring the evolution of, C. minuta in a subject.