Integrated Internal Standard Reagent for Cell Disruption
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Solution Overview
Problem
Current methods for disrupting cellular material often require external standards, which can introduce errors and are not effective in assessing the completeness of disruption and subsequent steps like sample preparation, extraction, and detection.
Innovation Solution
A reagent containing beads coated with reference cells is used for mechanical disruption, allowing an internal standard to be integrated for checking the completeness of disruption and subsequent steps, eliminating the need for a separate standard addition.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If external standards are used for checking disruption completeness, then the assessment of disruption and subsequent steps can be performed, but errors such as pipetting errors affecting only the sample material cannot be captured and the reliability is reduced
Solution Approach 1:
The patent combines the sample material and reference cells into a single reaction vessel, processing them together through all steps including disruption, extraction, and amplification. This merging ensures that any errors affecting the sample equally affect the reference cells, allowing detection of systematic errors while maintaining reliable assessment of disruption completeness.
Solution Approach 2:
The reference cells act as an intermediary standard that mediates between the sample material and the detection system. By being present throughout the entire process in the same reaction vessel, they provide a reliable reference point that captures systematic errors while enabling precise assessment of disruption completeness and subsequent processing steps.
2Reliability
If a separate standard addition step is used, then the checking concept can be implemented, but the device complexity and number of method steps increase
Solution Approach 1:
The patent merges the reference cells with the sample material in a single reaction vessel, eliminating the need for separate standard addition steps. The reference cells are added together with the sample at the beginning, and both are processed through disruption, extraction, and amplification in the same vessel, thereby reducing device complexity while maintaining reliability.
Solution Approach 2:
The reference cells serve multiple functions simultaneously: they act as a control for disruption completeness, a reference for extraction efficiency, and a monitor for amplification performance. This multi-functionality is achieved by having the reference cells present throughout all steps in the same reaction vessel, eliminating the need for separate checking steps.
3Measurement precision
If reference cells are added separately to the sample, then the internal standard concept can be implemented, but potential sources of error increase due to additional method steps
Solution Approach 1:
The patent combines the addition of reference cells and sample material into a single step, with both being added together to the reaction vessel. This merging eliminates additional method steps that could introduce errors, while maintaining the precision of internal standard assessment through co-processing of reference cells and sample through all subsequent steps.
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 ensures accurate assessment of cell disruption and subsequent processes, reducing errors and improving the reliability of nucleic acid analysis by using the coated beads as a stable internal standard.
Implementation Method 1
mechanical cell lysis, in which the cell wall is broken by mechanical stress
Implementation Method 2
the discovery of the polymerase chain reaction (PCR) in 1983, there has been development of a multiplicity of nucleic acid diagnostic methods
Data Source
AI summary
The present invention relates to a reagent for the disruption of cell material, containing an internal standard that is completely integrated into the reagent for control and evaluation of the completeness of disruption of the cell material and subsequent steps, comprising a step selected from sample preparation, extraction, enrichment, isolation, purification, reverse transcription, amplification and detection of the cell components obtained from the disrupted cells, or a combination of a plurality or all of these steps.