Multiplex Mastitis Test Strip for Rapid Pathogen Identification

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Solution Overview

Problem

Current methods for diagnosing bovine mastitis caused by E. coli, S. uberis, S. agalactiae, and S. aureus are time-consuming, require specialized equipment and skilled personnel, and cannot be performed in the field, leading to delayed targeted antibiotic therapy and the emergence of multi-resistant bacterial strains.

Innovation Solution

A rapid multiplex immunochromatographic test using monoclonal antibodies specific to ompA, EF-Tu, ADA, and FnBPA proteins from E. coli, S. uberis, S. agalactiae, and S. aureus, respectively, allowing for simultaneous identification of these pathogens in milk samples within 20 minutes without specialized equipment.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If culture-based, molecular, serology or spectrometric techniques are used to identify the etiological agent, then measurement precision and reliability are improved, but device complexity and ease of operation deteriorate, and loss of time increases

Engineering Contradiction:
Improveidentification accuracyVSAvoidtest duration
Core Design Contradiction:
Measurement precisionVSLoss of time

Solution Approach 1:

The test system is segmented into multiple independent detection channels, each targeting a specific bacterial pathogen (S. aureus, S. agalactiae, S. uberis, E. coli) with its own antibody-antigen interaction zone. This segmentation allows simultaneous detection of multiple pathogens in parallel, achieving both high identification accuracy and rapid results within 20 minutes.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

Multiple detection functions for different bacterial species are merged into a single immunochromatographic test device. The test strip integrates control line and multiple test lines that simultaneously detect various pathogens, combining what would traditionally require separate laboratory tests into one rapid field-testable unit.

Inventive Principle:
Principle #5Merging (Combining)

2Measurement precision

If culture-based, molecular, serology or spectrometric techniques are used to identify the etiological agent, then measurement precision is improved, but ease of operation and device complexity worsen

Engineering Contradiction:
Improveidentification accuracyVSAvoidoperational simplicity
Core Design Contradiction:
Measurement precisionVSEase of operation

Solution Approach 1:

The immunochromatographic test performs self-diagnosis through automatic capillary flow of the milk sample across the test strip. The system self-regulates the detection process without requiring external equipment or complex operational steps, enabling untrained personnel to conduct accurate pathogen identification simply by applying the sample and reading the results.

Inventive Principle:
Principle #25Self-service

Solution Approach 2:

Complex laboratory equipment (centrifuges, incubators, spectrometers) is replaced by a passive capillary-based immunochromatographic system. The test uses natural capillary action to move the sample and pre-immobilized antibodies to perform detection, eliminating the need for sophisticated mechanical or electronic equipment while maintaining identification accuracy.

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

3Loss of time

If rapid SCC estimation tests are used, then loss of time is reduced and ease of operation is improved, but measurement precision deteriorates because the etiological agent cannot be identified

Engineering Contradiction:
Improvetest durationVSAvoidpathogen identification accuracy
Core Design Contradiction:
Loss of timeVSMeasurement precision

Solution Approach 1:

The test strip is designed with multi-functionality, serving both as a rapid diagnostic tool and a specific pathogen identification system. It includes a control line for quality assurance and multiple test lines targeting different bacterial species, enabling simultaneous rapid detection and precise identification of the etiological agent in a single operation.

Inventive Principle:
Principle #6Universality (Multi-functionality)

Solution Approach 2:

The test changes the detection parameter from general somatic cell count estimation to specific antigen-antibody binding detection. By using pathogen-specific antibodies immobilized on the test strip, the system transforms the detection approach to identify specific bacterial antigens, achieving both speed and pathogen-level precision.

Inventive Principle:
Principle #35Parameter changes

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 test achieves high sensitivity, specificity, and accuracy of at least 75%, enabling on-site diagnosis by untrained personnel, thereby facilitating timely antibiotic therapy and reducing the risk of multi-resistant strains.

Implementation Method 1

A rapid multiplex immunochromatographic test using monoclonal antibodies specific to ompA, EF-Tu, ADA, and FnBPA proteins from E. coli, S. uberis, S. agalactiae, and S. aureus, respectively, allowing for simultaneous identification of these pathogens in milk samples

Methodology Applied
Scientific EffectAntigen-antibody binding:

Data Source

PatentEP4343328B1Diagnostic method and composition for the detection of mastitis caused by e. coli, s. uberis, s. agalactiae or s. aureus
Publication Date: 2026.03.11 JAGIELLONIAN UNIVERSITY
  • EP4343328B1 patent drawingFigure 1
  • EP4343328B1 patent drawingFigure 2~3
  • EP4343328B1 patent drawingFigure 4~5B

AI summary

A diagnostic method and composition are disclosed for detecting mastitis caused by E. coli, S. uberis, S. agalactiae or S. aureus, having at least 75% sensitivity, specificity and/or accuracy and useful for performing a test to detect mastitis in cows caused by the hereinabove pathogens easily and rapidly.