Bead Beating System With Dry Blocking Agent

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Solution Overview

Problem

Current methods for lysing cells and extracting nucleic acids from microorganisms in samples are complex and suffer from nucleic acid loss due to absorption on beads, and require dilution with lysis buffers, which limits detection accuracy and sensitivity.

Innovation Solution

The use of bead beating systems with integrated dry blocking agents that prevent nucleic acid absorption on beads, allowing for effective lysis and extraction without the need for pre-mixing with lysis buffers, utilizing chaotropic agents, detergents, nucleotides, and oligonucleotides to enhance nucleic acid recovery.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If lysis buffers are added to the sample fluid, then cell lysis and nucleic acid extraction are facilitated, but the sample fluid is diluted and DNA concentration decreases

Engineering Contradiction:
Improvenucleic acid extraction efficiencyVSAvoidDNA concentration
Core Design Contradiction:
ReliabilityVSQuantity of substance

Solution Approach 1:

The patent extracts the blocking agent function from the lysis buffer and incorporates it directly into the bead beating system. This allows the lysis buffer to be omitted, preventing sample dilution while maintaining nucleic acid extraction efficiency. The blocking agent is integrated into the beads or tube, providing its function independently of the lysis buffer.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The bead beating system is enhanced with multi-functionality by integrating blocking agents that simultaneously prevent nucleic acid adsorption to beads and facilitate efficient lysis. This multi-functional design eliminates the need for separate lysis buffer addition, maintaining both extraction efficiency and sample concentration.

Inventive Principle:
Principle #6Universality (Multi-functionality)

2Ease of manufacture

If bead beating is performed without blocking agents, then nucleic acid absorption on beads occurs, but the process is simpler

Engineering Contradiction:
Improveprocess simplicityVSAvoidnucleic acid loss
Core Design Contradiction:
Ease of manufactureVSLoss of substance

Solution Approach 1:

The blocking agent is incorporated into the bead beating system in advance, before the nucleic acid extraction process begins. This preliminary incorporation ensures that blocking sites are available from the start, preventing nucleic acid adsorption to beads during the beating process and subsequent handling.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The blocking agent acts as an intermediary substance that prevents direct interaction between nucleic acids and bead surfaces. By introducing this intermediary, the patent eliminates harmful adsorption while maintaining the mechanical lysis effectiveness of the bead beating process.

Inventive Principle:
Principle #24Intermediary (Mediator)

3Reliability

If more lysis buffer is used to prevent nucleic acid absorption, then absorption is reduced, but sample dilution increases

Engineering Contradiction:
Improvenucleic acid recoveryVSAvoidsample concentration
Core Design Contradiction:
ReliabilityVSQuantity of substance

Solution Approach 1:

The patent extracts the blocking agent function from the lysis buffer system and relocates it to the bead beating system. This separation allows blocking to occur without the dilution effect of adding large volumes of lysis buffer, achieving both high recovery and high concentration.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent changes the parameter of where blocking occurs - from occurring in solution (requiring buffer) to occurring at the bead surface interface. This parameter change allows blocking to happen with minimal or no buffer addition, preventing dilution while ensuring nucleic acid recovery.

Inventive Principle:
Principle #35Parameter changes

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 higher extraction rates of DNA and RNA, reducing sample dilution and nucleic acid loss, thereby improving detection sensitivity and accuracy in genetic testing.

Implementation Method 1

a plurality of particles movable relative to each other are introduced in the sample fluid and the sample fluid with the particles contained therein is oscillated such that the particles are capable of mechanically destroying cell walls of the microorganisms contained in the sample fluid

Methodology Applied
Scientific EffectMechanical force: Mechanical Force

Implementation Method 2

the use of bead beating in nucleic acid extraction methods results in nucleic acid loss due to absorption on the beads and that such loss can be prevented by the appropriate amount of blocking agent to block binding sites on the beads

Methodology Applied
Scientific EffectAdsorption: Adsorption

Data Source

PatentEP3408388B1Method for producing a lysate from cells contained in a liquid sample
Publication Date: 2021.10.20 SAFEGUARD BIOSYSTEMS HOLDINGS LTD
  • EP3408388B1 patent drawingFigure 1
  • EP3408388B1 patent drawingFigure 2~3
  • EP3408388B1 patent drawingFigure 4

AI summary

The present disclosure provides improved methods for bead beating and a bead beating system useful therefor. The bead beating system comprises a sample tube, beads, and a dry blocking agent, and methods for using the bead beating system to extract nucleic acids from cells containing the nucleic acids.