Two-Step Decontamination of Laboratory Consumables
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Solution Overview
Problem
Current methods for decontaminating laboratory consumables, such as tubes and filters, from residual nucleic acids are inadequate, leading to 'false positives' in PCR and TMA reactions, and existing sterilization techniques like Sterrad® process do not completely eliminate amplifiable nucleic acids, especially in complex devices with filters.
Innovation Solution
A two-step decontamination process involving treatment with gaseous ethylene oxide followed by liquid or gaseous hydrogen peroxide at specific concentrations and temperatures to effectively eliminate residual nucleic acids without damaging materials like polypropylene or PVDF filters.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional sterilization methods are used, then microbial contamination is prevented, but amplifiable residual nucleic acids remain on the surface
Solution Approach 1:
The sterilization process is divided into two distinct steps: first conventional sterilization to eliminate microbes, then a second treatment specifically targeting residual nucleic acids. This segmentation allows each step to optimize for its specific function without compromise.
Solution Approach 2:
The patent applies a preliminary action by treating the consumable with a nucleic acid-inactivating agent before the amplification reaction. This pre-treatment ensures that any residual nucleic acids are rendered non-amplifiable in advance, preventing false positives.
2Object-affected harmful factors
If ionizing radiation is used to eliminate residual nucleic acids, then amplification specificity improves, but material degradation occurs and regulatory constraints increase
Solution Approach 1:
The patent replaces the physical/chemical mechanism of ionizing radiation with a biochemical mechanism using nucleic acid-inactivating agents. These agents specifically target and inactivate nucleic acids through chemical modification without the broad destructive effects of radiation on materials.
Solution Approach 2:
The patent changes the approach from high-energy physical treatment to controlled chemical treatment at mild temperatures and pressures. This parameter change preserves material integrity while achieving the same goal of eliminating amplifiable nucleic acids.
3Adaptability or versatility
If low stringency conditions are used for amplification, then detection sensitivity for diverse sequences improves, but false positives from residual nucleic acids increase
Solution Approach 1:
The patent applies preliminary anti-action by inactivating residual nucleic acids before the amplification reaction. This pre-neutralization prevents them from interfering with the reaction, allowing low stringency conditions to be used without the risk of false positives from contaminating nucleic acids.
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
The combined treatment thoroughly removes residual nucleic acids, preventing undesirable amplifications and ensuring consumables are free from amplifiable DNA and RNA, even when initially contained in microorganisms, thereby reducing the risk of false positives and maintaining material integrity.
Implementation Method 1
Enzymes such as endonucleases or chemical products such as psoralen may be used in order to render the residual nucleic acid molecules non-amplifiable
Implementation Method 2
a second step of treatment with hydrogen peroxide in liquid phase at a concentration between 25% and 60% and heated to 50°C-70°C, or gaseous phase
Data Source
Figure 1
Figure 2A~2B
Figure 3A~3B
AI summary
The invention relates to a new process for treatment of residual nucleic acids present on the surface of laboratory consumables. This process combines two treatment phases: i) Treating with ethylene oxide in gaseous phase; then ii) Treating said surface with hydrogen peroxide in liquid phase or in gaseous phase; The effect of this treatment is to avoid the amplification of said residual nucleic acids, in particular during PGR or TMA reactions.