HbA1c Fluorescence Detection in Small Blood Samples
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Solution Overview
Problem
Conventional methods for determining HbA1c in small blood samples, such as those less than 200µl, face challenges in accuracy and reproducibility, and there is a need for patient-friendly methods and kits that allow individuals to measure HbA1c levels easily and affordably without medical assistance.
Innovation Solution
A method involving a fluorophore-binding technique where fluorescence measurements are taken at specific times after sample contact, and absorbance measurements are taken after adding a haemoglobin-binding agent, allowing for accurate determination of HbA1c relative to total haemoglobin concentration in small blood samples, using a kit-of-parts with a fluorophore, buffered liquid, and haemoglobin-binding agent.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional photometric or immunoassay methods are used to measure HbA1c, then the measurement can be performed with standard equipment, but the accuracy and reproducibility deteriorate when using small blood samples less than 200μl
Solution Approach 1:
The patent changes the detection parameter from conventional photometric absorbance to fluorescence emission intensity. This parameter change enables highly sensitive detection of HbA1c in small blood samples, achieving accurate measurement with less than 200μl blood volume while maintaining measurement precision
Solution Approach 2:
The patent replaces conventional mechanical/chemical separation methods (column-based borate affinity, HPLC) with a fluorescence-based detection system. This substitution eliminates the need for complex separation apparatus and enables accurate HbA1c measurement in small samples using a simplified fluorescent probe approach
2Reliability
If conventional HbA1c measurement methods are used, then established protocols are available, but reproducibility deteriorates with small sample volumes, failing to meet NGSP standards of less than 3.5% CV
Solution Approach 1:
By changing from absorbance measurement to fluorescence emission measurement, the patent achieves superior signal-to-noise ratio and detection sensitivity. This parameter change enables reproducible HbA1c measurement in small samples, meeting the stringent NGSP reproducibility criterion of less than 3.5% coefficient of variation
Solution Approach 2:
The patent introduces a fluorescent probe as an intermediary that selectively binds to HbA1c. This intermediary amplifies the detection signal through fluorescence emission, enabling reliable and reproducible measurement of HbA1c even in very small blood sample volumes
3Measurement precision
If professional laboratory equipment is used for HbA1c measurement, then accurate results can be obtained, but the ease of operation deteriorates requiring medical practitioner assistance
Solution Approach 1:
The patent enables patients to perform self-testing at home using simple equipment. The fluorescent probe method requires only basic fluorescence measurement capability, allowing patients to obtain accurate HbA1c results without visiting healthcare facilities or requiring professional laboratory equipment
Solution Approach 2:
By replacing complex laboratory instrumentation with a fluorescence-based system, the patent dramatically simplifies the measurement process. This substitution maintains measurement accuracy while enabling easy operation by patients themselves, eliminating the need for medical practitioner assistance
4Measurement precision
If conventional measurement methods are used, then standard procedures are available, but the device complexity increases requiring expensive equipment and facilities
Solution Approach 1:
The patent replaces complex mechanical separation systems (HPLC, column chromatography) with a fluorescence-based detection system. This substitution dramatically reduces device complexity, requiring only a fluorescence spectrometer or detector while maintaining high measurement precision for HbA1c
Solution Approach 2:
The patent extracts the essential detection function from complex laboratory equipment and implements it through a simplified fluorescence-based system. By taking out only the necessary detection capability and eliminating unnecessary separation and processing equipment, the patent achieves accurate HbA1c measurement with minimal device complexity
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 method provides improved accuracy and precision for measuring HbA1c in small blood samples, meeting the need for reliable and cost-effective patient-friendly testing, with results comparable to standard methods and meeting NGSP certification criteria for precision.
Implementation Method 1
measuring the fluorescence (FLT3,4) at the wavelength X of the first detection liquid at one or more time points within the time interval T3-T4, at which the change in fluorescence over time is >0
Implementation Method 2
measuring the transmission (TRT6) of the second detection liquid at approximately 570nm at time T6
Data Source
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AI summary
The present invention relates to methods and kits-of-parts for determining the quantity of HbA1c relative to the concentration of haemoglobin in a blood sample comprising less than 200µl, the method comprising the steps of adding the fluorophore to the sample and measuring the fluorescence at one or more time points within the time interval, at which the change in fluorescence over time is >0, followed by measurements of haemoglobin by adding a haemoglobin-binding agent and measuring the change in transmission at approximately 570nm and comparing the obtained results with an internal standard.