Liquid Culture Mutation Assay for Slow-Growing Microbes

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Solution Overview

Problem

Current methods for determining mutation accumulation in microbiology, such as the mutation-accumulation (MA) experiment, often require growth on agar plates, which does not accurately reflect the natural environment of many microorganisms and excludes slow-growing or unculturable microbes.

Innovation Solution

A method involving liquid culture systems where microorganisms are inoculated and incubated in a liquid medium, allowing for single-cell bottlenecks to be generated, thereby enabling mutation accumulation assays that more accurately reflect natural conditions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If agar plate growth is used for mutation accumulation experiments, then mutation rates can be estimated, but the method excludes slow-growing or unculturable microbes and does not accurately reflect natural environments

Engineering Contradiction:

Improveapplicability to diverse microorganismsVSAvoidaccuracy of mutation rate estimation

Core Design Contradiction:

Adaptability or versatilityVSReliability

Solution Approach 1:

The patent changes the physical state of the culture medium from solid (agar plate) to liquid form, fundamentally altering the growth parameters to accommodate slow-growing and unculturable microbes while maintaining the ability to perform single-cell bottlenecks and mutation accumulation assays

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

Instead of forcing microbes to grow on agar plates as traditionally done, the patent inverts the approach by using liquid culture systems that allow microbes to grow in their natural-like environment, thereby capturing diversity that was previously inaccessible

Inventive Principle:

Principle #13The other way round (Inversion)

2Ease of operation

If agar plates are used for culturing, then single-cell bottlenecks can be established, but the method creates an artificial environment that does not mimic natural conditions

Engineering Contradiction:

Improveability to perform single-cell bottlenecksVSAvoidrepresentativeness of natural environment

Core Design Contradiction:

Ease of operationVSMeasurement precision

Solution Approach 1:

The patent employs liquid culture systems where hydraulic principles allow for easy manipulation and distribution of single cells through liquid medium, enabling single-cell bottlenecks to be established while maintaining a natural-like aquatic environment that better mimics microbial habitats

Inventive Principle:

Principle #29Pneumatics and hydraulics

3Productivity

If traditional agar plate methods are used, then mutation accumulation can be measured, but slow-growing microbes cannot be detected

Engineering Contradiction:

Improvedetection speedVSAvoidrange of detectable microorganisms

Core Design Contradiction:

ProductivityVSAdaptability or versatility

Solution Approach 1:

The patent introduces dynamic liquid culture systems that can accommodate varying growth rates, allowing slow-growing microbes to be detected over extended periods while maintaining the productivity needed for mutation accumulation measurements through continuous monitoring and automated sampling

Inventive Principle:

Principle #15Dynamics

Data Source

PatentUS20250027074A1Methods for culturing microorganisms in a liquid culture system

Publication Date: 2025.01.23 THE ARIZONA BOARD OF REGENTS ON BEHALF OF THE UNIV OF ARIZONA

AI summary

The present invention provides a method of culturing microorganisms normally grown on a solid in liquid media, methods of use thereof to determine the presence or absence of the microorganism in a sample, and methods of use in a mutation accumulation assay.