Blood Analyzing Method for Piroplasm Detection

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Solution Overview

Problem

Current methods for detecting piroplasm-infected red blood cells in dogs and horses are not accurate enough, as they fail to effectively differentiate infected cells from uninfected ones using existing fluorescent dyes and detection techniques.

Innovation Solution

A method involving the use of a blood sample mixed with a fluorescent dye and a cationic surfactant, such as quaternary ammonium salts, to enhance membrane permeability and differentiate infected red blood cells through fluorescence and scattered light information analysis using flow cytometry.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If a fluorescent dye is used to stain nucleic acid in red blood cells, then fluorescence information can be obtained for detection, but the ability to accurately differentiate piroplasm-infected red blood cells from uninfected ones is insufficient

Engineering Contradiction:
Improvedetection accuracy of piroplasm-infected red blood cellsVSAvoiddifferentiation capability between infected and uninfected cells
Core Design Contradiction:
Measurement precisionVSReliability

Solution Approach 1:

A cationic surfactant is introduced as an intermediary substance that selectively interacts with piroplasm-infected red blood cells. The surfactant binds to the infected cells, creating a distinct complex that enhances fluorescence signal and enables reliable differentiation from uninfected cells through flow cytometry analysis.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The invention changes the chemical parameters of the staining system by combining fluorescent dyes with cationic surfactants. This parameter change modifies the fluorescence characteristics of infected cells, creating a distinguishable signal pattern that improves detection accuracy and differentiation capability simultaneously.

Inventive Principle:
Principle #35Parameter changes

2Measurement precision

If existing fluorescent staining methods are used, then nucleic acid can be visualized, but the measurement precision for detecting piroplasm-infected red blood cells remains inadequate

Engineering Contradiction:
Improveparasitemia detection precisionVSAvoidinformation about infected cell characteristics
Core Design Contradiction:
Measurement precisionVSLoss of information

Solution Approach 1:

The invention utilizes fluorescence emission (optical color change) by combining specific fluorescent dyes with cationic surfactants. The surfactant-fluorophore complex produces enhanced and distinct fluorescence signals that provide rich information about infected cell characteristics, enabling precise parasitemia measurement without information loss.

Inventive Principle:
Principle #32Color 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 allows for precise detection and counting of piroplasm-infected red blood cells, improving the accuracy of piroplasm infection diagnosis in animals.

Implementation Method 1

a cationic surfactant, such as quaternary ammonium salts, to enhance membrane permeability

Methodology Applied
Scientific EffectMembrane permeability enhancement: Surfactant

Implementation Method 2

a fluorescent dye that stains nucleic acid... acquiring fluorescence information by irradiating with light a measurement sample

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Implementation Method 3

acquiring scattered light information... detecting the piroplasm-infected red blood cell in the blood sample based on the fluorescence information and the scattered light information

Methodology Applied
Scientific EffectLight scattering: Scattering

Data Source

PatentEP3842783B1Blood analyzing method
Publication Date: 2024.01.10 SYSMEX CORP
  • EP3842783B1 patent drawingFigure 1
  • EP3842783B1 patent drawingFigure 2
  • EP3842783B1 patent drawingFigure 3~4

AI summary

Disclosed is a blood analyzing method comprising: acquiring fluorescence information by irradiating with light a measurement sample prepared by mixing a blood sample collected from a living organism, a fluorescent dye that stains nucleic acid, and a cationic surfactant; and detecting a piroplasm-infected red blood cell in the blood sample based on the fluorescence information.