Eukaryotic Nucleic Acid Isolation from Stool Samples
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Solution Overview
Problem
Current methods for diagnosing gastrointestinal disorders, such as gastrointestinal cancer and inflammatory bowel disease, are invasive, time-consuming, and costly, and face challenges in distinguishing eukaryotic nucleic acids from the high bacterial load in stool samples, leading to unfavorable signal-to-noise ratios and degradation of eukaryotic signals.
Innovation Solution
A method involving the separation of eukaryotic cells from bacterial cells in stool samples using a buffer, surfactant, and ribonuclease inhibitor, followed by chaotropic agent treatment, to isolate eukaryotic nucleic acids, which can be analyzed for biomarkers using techniques like microarray sequencing and PCR, enabling the detection of gastrointestinal disorders.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If standard diagnostic methods like colonoscopy are used, then diagnostic accuracy is improved, but patient comfort and cost deteriorate due to invasiveness and high cost
Solution Approach 1:
The patent extracts eukaryotic nucleic acids from the complex stool sample matrix by separating them from bacterial cells through density gradient centrifugation. This extraction allows diagnostic analysis to be performed on isolated eukaryotic material rather than the entire stool sample, enabling non-invasive testing while maintaining diagnostic accuracy
Solution Approach 2:
The patent uses eukaryotic nucleic acids as an intermediary biomarker that can be detected in stool samples to indirectly diagnose gastrointestinal disorders. This intermediary approach allows diagnosis without direct visualization or intervention in the gastrointestinal tract, improving patient comfort while maintaining diagnostic capability
2Measurement precision
If standard diagnostic methods like colonoscopy are used, then diagnostic accuracy is improved, but time consumption and cost deteriorate
Solution Approach 1:
The patent performs preliminary isolation of eukaryotic nucleic acids from stool samples using a standardized protocol involving buffer treatment, centrifugation, and purification. This preliminary preparation creates ready-to-analyze samples that can be processed through various detection methods more quickly than traditional diagnostic procedures
Solution Approach 2:
The patent creates a copy or representation of the gastrointestinal disease state through detection of eukaryotic nucleic acid biomarkers in stool samples. This molecular copy allows diagnosis without requiring the time-consuming procedure of direct visualization through colonoscopy
3Ease of operation
If eukaryotic nucleic acids are isolated from stool samples, then non-invasive diagnosis is enabled, but signal-to-noise ratio deteriorates due to high bacterial load
Solution Approach 1:
The patent segments the complex stool sample into distinct components based on density, separating eukaryotic cells from bacterial cells through density gradient centrifugation. This segmentation isolates the eukaryotic nucleic acid signal from the overwhelming bacterial background, improving signal-to-noise ratio while maintaining the non-invasive nature of the test
Solution Approach 2:
The patent extracts eukaryotic nucleic acids specifically from the stool sample matrix by separating them from bacterial cells. This extraction process removes the bacterial noise while retaining the diagnostic eukaryotic signal, enabling non-invasive diagnosis with improved signal-to-noise ratio
4Ease of operation
If eukaryotic nucleic acids are isolated from stool samples, then non-invasive diagnosis is enabled, but nucleic acid degradation worsens the reliability of detection
Solution Approach 1:
The patent performs preliminary isolation of eukaryotic nucleic acids from stool samples using a standardized protocol that includes buffer treatment, centrifugation, and purification steps designed to protect nucleic acids from degradation. This preliminary preparation ensures nucleic acid stability while maintaining the non-invasive advantage of stool-based testing
Solution Approach 2:
The patent uses eukaryotic nucleic acids as a stable intermediary biomarker that can be isolated and protected from degradation during the diagnostic process. By focusing on isolated nucleic acids rather than whole cells or tissues, the method maintains reliability while enabling non-invasive diagnosis
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
This method effectively isolates eukaryotic nucleic acids, allowing for the detection of biomarkers indicative of gastrointestinal disorders, providing a non-invasive and cost-effective diagnostic approach for both humans and animals.
Implementation Method 1
separating the suspension into a portion enriched for eukaryotic cells and a portion enriched for bacterial cells
Implementation Method 2
adding a chaotropic agent and optionally a surfactant to the portion enriched for eukaryotic cells to form a lysate
Implementation Method 3
fractioning the lysate into a cell debris layer, a layer comprising eukaryotic nucleic acids and a lipid layer
Data Source
Figure 1(a)~1(b)
Figure 2
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AI summary
Provided herein are materials and methods for isolation of eukaryotic nucleic acid from a human or non-human animal stool sample. Also provided are methods of analysis of eukaryotic biomarkers present in a human or non-human animal stool sample.