Beta-Lactamase Enzyme Addition for Rapid Antibiotic Detection in Milk

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Solution Overview

Problem

Current microbial growth inhibition tests for detecting antibiotics in samples like milk are lengthy, often taking 2.5 to 4.5 hours, which delays processing and are sensitive to concentration changes when methods to reduce duration are implemented, such as using higher test organism concentrations or multiple indicators, leading to decreased sensitivity and increased costs.

Innovation Solution

Incorporating beta-lactamase into the test organism, sample, or both, to reduce the test duration while maintaining sensitivity, by adding it before or during the test, and using a suitable indicator and gelling agent in a test medium that can be in liquid, solid, or gel form, with specific concentrations of beta-lactamase and indicators.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Loss of time

If the test duration is reduced by using higher test organism concentrations or multiple indicators, then the test duration decreases, but the sensitivity of the test decreases and costs increase

Engineering Contradiction:
Improvetest durationVSAvoidtest sensitivity
Core Design Contradiction:
Loss of timeVSMeasurement precision

Solution Approach 1:

A beta-lactamase enzyme is introduced as an intermediary substance that specifically degrades beta-lactam antibiotics in the sample before the test organism is added. This pre-treatment step removes the inhibitory effect of beta-lactam antibiotics, allowing the test to proceed normally and achieve both reduced duration and maintained sensitivity without requiring higher organism concentrations or multiple indicators

Inventive Principle:
Principle #24Intermediary (Mediator)

2Loss of time

If the test duration is reduced by using higher test organism concentrations, then the test duration decreases, but the cost of the test increases

Engineering Contradiction:
Improvetest durationVSAvoidtest cost
Core Design Contradiction:
Loss of timeVSQuantity of substance

Solution Approach 1:

The beta-lactamase enzyme acts as a cost-effective intermediary that enables rapid testing by pre-degrading beta-lactam antibiotics. This approach avoids the need to use large quantities of expensive test organisms, achieving both time reduction and cost control through a single enzyme addition step

Inventive Principle:
Principle #24Intermediary (Mediator)

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 addition of beta-lactamase significantly shortens the test duration without compromising sensitivity, allowing for quicker detection of antibiotics in milk samples, making the test more efficient and cost-effective.

Implementation Method 1

a beta-lactamase is added to the test organism, the sample or both and the duration of the test is reduced by the addition of the beta-lactamase

Methodology Applied
Scientific EffectBeta-lactamase enzymatic degradation: Enzyme

Implementation Method 2

growing the test organism, detecting the amount of growth of the test organism

Methodology Applied
Scientific EffectMicrobial growth: Fermentation

Data Source

PatentUS9523112B2Method for the determination of the presence of an antibiotic in a fluid
Publication Date: 2016.12.20 DSM IP ASSETS BV

AI summary

The present invention provides a method and test for the determination of the presence or absence of an antibiotic in a sample such as milk.