Microbial DNA Isolation via Filtration Matrix
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Solution Overview
Problem
Current methods for isolating nucleic acids from microbial cells in mixed samples with higher eukaryotic cells or tissues are laborious, time-consuming, and require significant manual intervention, often hindered by interference from excess eukaryotic DNA.
Innovation Solution
A method involving filtration of a liquid sample through a nucleic acid-binding glass or silica matrix, followed by lysis and binding of nucleic acids on the matrix using cations, allowing for efficient and automated purification of microbial DNA without precipitation, enabling high purity and reduced processing time.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If centrifugation-based methods are used for DNA purification, then DNA can be isolated from mixed samples, but the processing time is long and manual intervention is required
Solution Approach 1:
The invention combines cell filtration and DNA binding onto a single filter matrix, merging two separate operations (filtration and purification) into one integrated system. This eliminates the need for separate centrifugation steps and reduces manual intervention, directly addressing the contradiction between processing time and ease of operation.
Solution Approach 2:
The filter matrix serves multiple functions simultaneously: it acts as a physical barrier to retain cells, provides a surface for DNA binding, and enables automated processing. This multi-functionality reduces the number of steps required and minimizes manual intervention while maintaining efficient DNA isolation.
2Measurement precision
If conventional DNA purification kits are used, then DNA can be isolated, but excess eukaryotic DNA interferes with detection of microbial DNA
Solution Approach 1:
The invention selectively extracts microbial DNA from mixed samples containing both microbial and eukaryotic cells. By using specific lysis conditions that target microbial cells while preserving eukaryotic cell integrity, the method extracts only the desired microbial DNA and leaves eukaryotic DNA behind, eliminating interference in detection.
Solution Approach 2:
The method applies different lysis conditions to different cell types within the same sample. Microbial cells are lysed under specific conditions that release their DNA, while eukaryotic cells remain intact. This localized differentiation in treatment quality enables selective DNA extraction and improves detection accuracy.
3Measurement precision
If sophisticated methods with immobilized proteins are used to separate human DNA from bacterial DNA, then detection accuracy improves, but the device complexity and cost increase
Solution Approach 1:
The invention uses a simple, disposable filter matrix instead of complex immobilized protein systems. The filter matrix is a inexpensive, single-use component that provides sufficient separation capability without requiring sophisticated equipment or complex methodologies, thus reducing device complexity while maintaining adequate detection accuracy.
Solution Approach 2:
The method achieves DNA separation by changing physical and chemical parameters (pH, ionic strength, lysis conditions) rather than using complex immobilized protein systems. This parameter-based approach simplifies the methodology and reduces device complexity while still enabling effective separation of microbial DNA from eukaryotic DNA.
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 achieves rapid and efficient isolation of nucleic acids with high purity, reducing processing time by half compared to centrifugation-based methods and facilitating automation, thereby overcoming the limitations of existing techniques.
Implementation Method 1
binding the nucleic acids to the matrix by addition of monovalent and/or multivalent cations
Implementation Method 2
filtering a liquid sample through a nucleic acid-binding glass or silica matrix which has a pore size small enough to retain microbial cells
Implementation Method 3
lysis of the microbial cells on the matrix by a combination of a lytic enzyme, a salt solution and a detergent
Data Source
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AI summary
The present invention relates to a new method for enriching and/or isolating nucleic acids from microbial cells which comprises filtering a liquid sample through a nucleic acid-binding matrix which has a pore size small enough to retain microbial cells, lysing the microbial cells on the matrix to release the nucleic acids from the microbial cells, binding the nucleic acids to the matrix and subsequently eluting the DNA. The invention also relates to a method for enriching and/or isolating nucleic acids from microbial cells which are present in a liquid sample that comprises microbial cells and higher eukaryotic cells and/or tissues. The invention also provides a cartridge for carrying out the methods of the invention. Finally, the invention relates to kits for carrying out the methods of the invention.