Fluorescent Spore Indicator for Rapid Oxidative Sterilization Checks

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Solution Overview

Problem

Existing biological indicators for sterilization processes are costly, time-consuming, and unreliable due to their reliance on complex procedures and enzymatic reactions, which are sensitive to environmental conditions and enzyme stability, leading to potential inaccuracies.

Innovation Solution

A self-contained biological indicator using a fluorescence intensity test and colorimetric test after incubation, employing microbial spores and fluorescent sensor proteins that detect structural changes in response to oxidative sterilization, independent of catalytic activity, within a flexible container.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If complex enzymatic reactions and genetically engineered microorganisms are used for biological indicators, then measurement precision may be improved, but device complexity and cost increase significantly

Engineering Contradiction:
Improvesterilization efficacy determination accuracyVSAvoidbiological indicator system complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent extracts and eliminates the complex enzymatic reaction components and genetically engineered microorganisms from the biological indicator system. Instead, it uses simple microbial spores with inherent catalase enzyme that naturally produces oxygen bubbles when exposed to hydrogen peroxide, providing a straightforward visual indicator without requiring complex genetic engineering or multiple enzymatic steps.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent employs simple, inexpensive microbial spores that can be easily prepared and discarded after use, replacing expensive and complex genetically engineered microorganisms. The spores serve as single-use indicators that provide reliable results without requiring sophisticated equipment or complex handling procedures.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

2Loss of time

If enzymatic activity tests are performed immediately after sterilization, then time consumption is reduced, but reliability decreases due to enzyme instability and sensitivity to environmental conditions

Engineering Contradiction:
Improvesterilization verification timeVSAvoidsterilization efficacy determination reliability
Core Design Contradiction:
Loss of timeVSReliability

Solution Approach 1:

The patent utilizes the natural catalase enzyme present in microbial spores that automatically reacts with residual hydrogen peroxide through inherent catalytic activity. This self-service mechanism eliminates the need for external enzymatic reactions or complex testing procedures, providing reliable and immediate results based on the spores' own biochemical properties without being affected by environmental variations.

Inventive Principle:
Principle #25Self-service

3Measurement precision

If specialized equipment and lengthy incubation procedures are used, then measurement precision is improved, but productivity and ease of operation deteriorate

Engineering Contradiction:
Improvesterilization outcome assessment accuracyVSAvoidsterilization process throughput
Core Design Contradiction:
Measurement precisionVSProductivity

Solution Approach 1:

The patent replaces complex mechanical and equipment-based detection systems with a simple visual observation method. Instead of using specialized incubators, spectrophotometers, or other sophisticated equipment, the invention relies on direct visual detection of oxygen bubble formation, which can be observed with the naked eye, thereby eliminating the need for expensive equipment and lengthy incubation periods while maintaining measurement precision.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

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

Provides immediate and reliable results, reducing costs and time requirements by avoiding lengthy incubation and specialized equipment, while ensuring accurate determination of sterilization efficacy through structural protein changes.

Implementation Method 1

The at least one fluorescent sensor protein is capable of yielding an optically detectable signal, i.e., a fluorescence signal, when the at least one sensor protein is not in a denatured state due to the oxidative sterilization process

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Implementation Method 2

a sterilization process employing hydrogen peroxide

Methodology Applied
Scientific EffectOxidation: Oxidation

Data Source

PatentEP4363602B1Biological indicator for determining the efficacy of an oxidative sterilization process and methods of use
Publication Date: 2025.11.26 TERRAGENE LLC
  • EP4363602B1 patent drawingFigure 1
  • EP4363602B1 patent drawingFigure 2
  • EP4363602B1 patent drawingFigure 3

AI summary

A biological indicator for determining the efficacy of an oxidative sterilization process, and its methods of use. The biological indicator comprises a set of microbial spores, at least one fluorescent sensor protein, and a culture medium, the fluorescent sensor protein being capable of yielding an optically detectable signal when the fluorescent sensor protein is not in a denatured state due to the oxidative sterilization process, and a different optically detectable signal when the fluorescent sensor protein is in a denatured state after the oxidative sterilization process.