Immunoenzymatic LH Detection Kit for Mammals

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Solution Overview

Problem

Current methods for detecting the preovulatory LH peak in mammals are not reliable, prone to false positives and negatives, and are not adaptable for use in various biological samples or animal breeding conditions, leading to inaccurate prediction of ovulation time.

Innovation Solution

A method involving an anti-LH antibody attached to a test surface, treated with a buffer solution and then contacted with a biological sample, followed by enzyme-conjugate and substrate solutions to achieve specific and sensitive detection of the LH peak, allowing for visual interpretation without specialized equipment.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If urinary tests are used for LH detection, then the test can be applied to human females, but the test is difficult to apply to mammals such as pigs, sheep, cattle, goats and requires additional costs for urine collection

Engineering Contradiction:
Improveadaptability to different mammalsVSAvoidease of sample collection
Core Design Contradiction:
Adaptability or versatilityVSEase of operation

Solution Approach 1:

The test strip is designed to detect LH in multiple biological matrices (urine, blood, saliva, vaginal mucus) using the same immunoenzymatic principle, making it universally applicable across different mammal species without requiring species-specific test designs. The antibody-based detection mechanism works across species boundaries for LH detection.

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

Solution Approach 2:

The patent uses saliva as an intermediary sample type that can be easily collected from various mammals without requiring invasive procedures or specialized equipment. Saliva collection tubes with stabilizing agents serve as a universal sampling method that bridges the gap between urinary tests (human-specific) and blood tests (invasive but species-independent).

Inventive Principle:
Principle #24Intermediary (Mediator)

2Ease of operation

If lateral flow tests are used for LH detection, then the test can detect presence or absence of LH, but the interpretation of results is ambiguous and presents significant number of false negatives or false positives

Engineering Contradiction:
Improvesimplicity of test formatVSAvoidaccuracy of result interpretation
Core Design Contradiction:
Ease of operationVSReliability

Solution Approach 1:

The test uses colorimetric changes through enzyme-substrate reactions (TMB, DAB, CN) that produce distinct, unambiguous color signals indicating LH detection. The color intensity correlates with LH concentration, providing clear visual differentiation between positive and negative results without the ambiguous band interpretation problems of lateral flow tests.

Inventive Principle:
Principle #32Color changes

Solution Approach 2:

The patent replaces the mechanical capillary action-based lateral flow mechanism with an immunoenzymatic system where antibodies bound to LH are detected through enzyme-catalyzed color reactions. This substitution eliminates the ambiguous visual interpretation of flow bands while maintaining ease of use through simple color reading.

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

The method provides near-quantitative detection of the LH peak with high specificity and sensitivity, reducing false positives and negatives, and can be used with various biological samples, including blood, urine, and saliva, facilitating precise prediction of ovulation.

Implementation Method 1

contacting the test surface rinsed in step (d) with a conjugate buffer solution comprising an anti-LH antibody coupled to an enzyme

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

bringing the test surface into contact with a solution comprising a substrate of said enzyme, said substrate being chosen from the group comprising 3,3'3,3',5,5'-tetramethylbenzidine (TMB membrane), 4-Chloro-1-naphtho/3,3'-diaminobenzidine tetrahydrochloride (CN/DAB), 3-amino-9-ethylcarbazole (AEC)

Methodology Applied
Scientific EffectColorimetric reaction: Chemical Bonding

Implementation Method 3

attach an anti-LH antibody to a test surface; bringing said surface obtained in step (b) into contact with a biological sample

Methodology Applied
Scientific EffectAntigen-antibody binding: Chemical Bonding

Implementation Method 4

contact the test surface on which said anti-LH antibody is fixed with a buffer solution comprising 5 to 50% by volume of fetal calf serum

Methodology Applied
Scientific EffectBlocking: Adsorption

Data Source

PatentEP2836839B1Method and kit for detecting the LH preovulatory surge
Publication Date: 2017.10.04 REPROPHARM VET
  • EP2836839B1 patent drawingFigure 1A~1B
  • EP2836839B1 patent drawingFigure 2A~2B
  • EP2836839B1 patent drawingFigure 3A~4

AI summary

The present invention relates to a method for detecting the LH preovulatory surge in a biological sample from mammals and a kit for implementing the detection method. The present invention can be applied, in particular, to the veterinary and medical fields.