Selective Cell Lysis Using Ionic Surfactants
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Solution Overview
Problem
Current methods for detecting pathogens in blood samples are hindered by the presence of high amounts of eukaryotic DNA, which interferes with PCR reactions, increases sample viscosity, and forms aggregates, making it challenging to detect minute amounts of bacterial DNA without extensive purification and dilution.
Innovation Solution
A method involving the use of an ionic surfactant, such as sodium dodecyl sulfate, in combination with an alkaline buffer at a pH of 9.0 or higher to selectively lyse eukaryotic cells, allowing for the retention of microorganisms and subsequent filtration, eliminating the need for enzymatic DNA degradation and reducing sample viscosity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If large volumes of blood are processed to detect minute amounts of bacteria, then detection sensitivity is improved, but sample viscosity increases and manipulation time becomes unacceptable
Solution Approach 1:
The patent extracts and removes eukaryotic DNA from the blood sample using selective lysis with ionic surfactants and alkaline conditions. This extraction eliminates the interfering substance (eukaryotic DNA) while preserving bacterial DNA, allowing sensitive detection without requiring extensive dilution or time-consuming manipulation steps
Solution Approach 2:
The patent changes the chemical parameters of the sample by adjusting pH to alkaline conditions (pH 9.0 or higher) and introducing ionic surfactants. These parameter changes enable selective lysis of eukaryotic cells while preserving bacterial integrity, thereby improving detection sensitivity without increasing manipulation time
2Difficulty of detecting and measuring
If eukaryotic cells are lysed to access intracellular components, then analytical capability is improved, but sample viscosity increases and filtration is prevented
Solution Approach 1:
The patent applies local quality by using ionic surfactants that selectively interact with eukaryotic cell membranes under alkaline conditions. The surfactants locally disrupt eukaryotic membranes while leaving bacterial cells intact, enabling analytical access to eukaryotic components without creating the aggregation and viscosity problems that would prevent filtration
Solution Approach 2:
The patent introduces ionic surfactants as intermediaries that mediate the lysis process. These surfactants act as intermediaries between the alkaline environment and the eukaryotic cell membranes, enabling controlled lysis that improves analytical capability while maintaining sample properties suitable for filtration
3Measurement precision
If DNA purification is performed to remove eukaryotic DNA, then detection accuracy is improved, but process complexity increases
Solution Approach 1:
The patent performs preliminary action by conducting selective lysis of eukaryotic cells before the PCR detection step. This preliminary removal of eukaryotic DNA through ionic surfactant treatment and alkaline conditions simplifies the overall process by eliminating the need for complex DNA purification protocols, thereby improving detection accuracy while reducing process complexity
Solution Approach 2:
The patent converts the harmful effect of high eukaryotic DNA content into a benefit by using the same DNA-rich environment to drive the selective lysis reaction. The high concentration of eukaryotic DNA, which initially causes interference, becomes the target of selective lysis, and the resulting debris actually aids in the separation and detection of bacterial 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 approach enables the processing of larger blood volumes with minimal dilution, maintaining the integrity of microorganisms, and facilitates automated manipulation, allowing for effective detection of pathogens without the complexity of prior art methods.
Implementation Method 1
a method involving the use of an ionic surfactant, such as sodium dodecyl sulfate, in combination with an alkaline buffer at a pH of 9.0 or higher to selectively lyse eukaryotic cells
Implementation Method 2
adding a buffer having a pH of about 9.0 or more, preferably a pH of about 9.5 or more and an ionic surfactant to the sample to obtain a solution having a pH of 9.0 or more
Data Source
Figure 1~2
Figure 3~4
AI summary
The present invention discloses methods, kits-of-parts, and devices for the selective lysis of eukaryotic cells in a sample comprising micro-organisms such as bacteria unicellular fungi. The selective lysis is obtained by incubating the sample in an ionic surfactant under alkaline conditions.