Avian Biliverdin Assay Enzymatic Conversion Spectrophotometry

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Solution Overview

Problem

Current biochemical tests for liver disease in birds and reptiles are limited, as they rely on bilirubin measurement, which is not applicable due to the absence of bilirubin in these species, necessitating a method to quantify biliverdin for effective diagnosis.

Innovation Solution

A method involving the use of biliverdin reductase to convert biliverdin to bilirubin, allowing for the measurement of biliverdin concentration in avian and reptilian samples by detecting changes in absorbance at specific wavelengths, with results compared to standard concentration curves.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If bilirubin measurement is used for diagnosing liver disease in birds and reptiles, then the diagnostic method is simple and widely applicable, but the measurement is not applicable because bilirubin is not produced in these species

Engineering Contradiction:

Improveapplicability of bilirubin measurementVSAvoiddiagnostic accuracy

Core Design Contradiction:

Adaptability or versatilityVSReliability

Solution Approach 1:

The patent changes the measurement parameter from bilirubin concentration to biliverdin concentration. Since birds and reptiles lack biliverdin reductase and therefore do not produce bilirubin, the invention measures biliverdin instead, which is the actual waste product of hemoglobin destruction in these species. This parameter change makes the diagnostic test applicable to avian and reptilian physiology.

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent uses biliverdin as an intermediary substance to indirectly assess liver disease. Instead of measuring bilirubin directly (which is not produced in these species), the invention measures biliverdin, which serves as an intermediate marker that reflects the same pathophysiological processes (hemoglobin destruction and hepatic function) but is actually present in avian and reptilian systems.

Inventive Principle:

Principle #24Intermediary (Mediator)

2Reliability

If biliverdin measurement is implemented in birds and reptiles, then diagnostic accuracy for liver disease is improved, but the assay complexity increases compared to standard bilirubin tests

Engineering Contradiction:

Improvediagnostic accuracyVSAvoidassay complexity

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent replaces the need for complex chromatographic separation methods with a spectrophotometric assay. Instead of using HPLC or other complex separation techniques to measure biliverdin, the invention uses a simplified spectrophotometric method that measures absorbance changes at specific wavelengths (325-500 nm or 500-750 nm) after enzymatic conversion, making the test more accessible to routine laboratory settings.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent optimizes the measurement wavelength parameters to simplify the assay. By measuring absorbance changes at specific wavelength ranges (325-500 nm or 500-750 nm), the invention creates a standardized parameter set that can be easily implemented in routine laboratories without requiring complex instrumentation or multiple measurement steps.

Inventive Principle:

Principle #35Parameter changes

3Ease of operation

If biliverdin reductase is used to convert biliverdin to bilirubin for measurement, then the measurement can be performed using standard spectrophotometric methods, but the enzyme conversion process adds steps to the assay

Engineering Contradiction:

Improveease of measurementVSAvoidassay steps

Core Design Contradiction:

Ease of operationVSDevice complexity

Solution Approach 1:

The patent uses biliverdin reductase as an intermediary enzyme to facilitate the measurement. The enzyme converts biliverdin to bilirubin, which has different absorbance characteristics that are easier to measure standardly. This enzymatic intermediary step simplifies the measurement process by transforming the target analyte into a more measurable form using conventional spectrophotometric techniques.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent changes the chemical state of biliverdin through enzymatic conversion to bilirubin, which alters the absorbance parameters and enables measurement using standard spectrophotometric methods. This parameter change in the chemical form of the analyte simplifies the operational aspects of the assay by allowing use of conventional measurement equipment and procedures.

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

This approach enables accurate quantitation of biliverdin levels, aiding in the diagnosis and differentiation of intra- and extrahepatic diseases in birds and reptiles, providing a more effective diagnostic tool than existing methods.

Implementation Method 1

contacting the sample with biliverdin reductase

Methodology Applied

Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

conversion of biliverdin to bilirubin

Methodology Applied

Scientific EffectReduction reaction: Reduction

Implementation Method 3

measuring a change in absorbance at about 325 nm to about 500 nm

Methodology Applied

Scientific EffectAbsorbance spectroscopy: Absorption Spectroscopy

Data Source

PatentUS7785822B2Assays for the detection of biliverdin in birds and reptiles

Publication Date: 2010.08.31 UNIVERSITY OF GEORGIA RESEARCH FOUNDATION INC

AI summary

The present invention provides methods for the detection of biliverdin in birds (avian species) and reptiles.