Genetically-Modified Spore Indicator for Rapid Sterilization Assessment
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Solution Overview
Problem
Conventional biological sterilization indicators require extended time to determine the efficacy of sterilization processes, as they rely on the growth of test microorganisms in a liquid medium, which can take up to 24 hours, hindering rapid assessment of sterilization effectiveness.
Innovation Solution
A self-contained sterilization process biological indicator using genetically-modified test microorganisms that produce a chimeric protein with enhanced enzyme activity, allowing for rapid detection of spore viability by reacting with an enzyme substrate to form a detectable product, significantly reducing the time needed to assess sterilization efficacy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional biological indicators use test microorganisms grown in liquid medium, then reliable detection of spore viability is achieved, but the detection time is extended to up to 24 hours
Solution Approach 1:
The patent replaces the conventional mechanical/biological growth-based detection system with an enzyme-catalyzed chemical reaction system. Instead of waiting for microorganism growth in liquid medium, the invention uses enzyme substrates that rapidly react with enzymes from viable spores to produce detectable signals (color change, fluorescence, or chemiluminescence), reducing detection time from 24 hours to minutes while maintaining reliability
Solution Approach 2:
The invention changes the detection parameter from measuring microbial growth (which takes time) to measuring enzyme activity through chemical reactions. By using enzyme substrates that produce immediate detectable products upon enzymatic cleavage or modification, the system transforms a slow biological process into a rapid chemical detection process
2Measurement precision
If conventional biological indicators require incubation in liquid medium, then accurate assessment of sterilization efficacy is achieved, but the process complexity and time consumption increase
Solution Approach 1:
The patent extracts the essential detection function from the complex liquid medium incubation process. By isolating and utilizing only the enzyme activity present in viable spores, the invention eliminates the need for complex nutrient media, incubation equipment, and extended observation periods, simplifying the overall detection system while preserving accuracy
Solution Approach 2:
The invention introduces enzyme substrates as intermediaries that bridge the gap between spore viability and detectable signal. These substrates act as mediators that rapidly convert enzymatic activity into measurable outputs (colorimetric, fluorometric, or chemiluminescent signals), providing accurate assessment without requiring complex biological culture systems
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 use of genetically-modified test microorganisms with chimeric proteins enables rapid detection of sterilization effectiveness, reducing the time required to determine spore viability from potentially days to less than an hour, providing a quicker and more efficient assessment of sterilization processes.
Implementation Method 1
the chimeric protein comprises a first segment and a second segment that is contiguous with the first segment. The first segment can comprise at least a portion of a first polypeptide that is normally found in spores. The second segment can comprise a second polypeptide having a detectable enzymatic activity.
Implementation Method 2
an analysis of the fluorescence intensity due to a fluorescent product of an enzyme reaction serves to determine whether the sterilization process was successful
Data Source
AI summary
An article for assessing efficacy of a sterilization process is provided. The article includes a housing, a plurality of genetically-modified spore-forming test microorganisms disposed in the housing, a liquid medium disposed in an openable container that is disposed in or attached to the housing, and an enzyme substrate disposed in the housing or in the openable container. The genetically-modified test microorganism comprises a functional fusion gene that encodes a non-naturally occurring chimeric protein, the chimeric protein comprising a first segment and a second segment that is contiguous with the first segment. The first segment comprises at least a portion of a first polypeptide that is normally found in spores and the second segment comprises a second polypeptide having a detectable enzymatic activity. The enzyme substrate is capable of reacting with the detectable enzyme activity to form a detectable product. A method of using the test microorganisms is also provided.