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Fully Automated Centrifugal Microfluidic Device for Ultrasensitive Protein Detection from Whole Blood
Published on: April 16, 2016
Development of a canine blood C-reactive protein-measuring device using a flow-type immunosensor
Tomoko Kubota1, Norio Tateishi1, Hideki Toita2
1Kyoto Bio Laboratory, Seeds Tec Co., Ltd., Kyodai Katsura Venture Plaza South Building 2113, Goryo-Ohara 1-39, Sakyo-ku, Kyoto, 6158245, Japan.
Insights
A novel flow-type immunosensor (FIS) achieves performance comparable to automated analyzers for measuring canine C-reactive protein (CRP). This fluorescence-linked immunosorbent assay technology offers accurate and reliable CRP quantification in canine plasma.
Area of Science:
- Biomedical Engineering
- Analytical Chemistry
- Veterinary Diagnostics
Background:
- Automated analyzers and immunoturbidimetric reagents are standard for C-reactive protein (CRP) measurement.
- Developing alternative, high-performance diagnostic tools is crucial for veterinary medicine.
- Flow-type immunosensors (FIS) offer potential for rapid and sensitive immunoassays.
Purpose of the Study:
- To construct a flow-type immunosensor (FIS) for canine C-reactive protein (CRP) measurement.
- To evaluate the analytical performance of the developed FIS.
- To compare the FIS performance against a standard automated analyzer method.
Main Methods:
- Construction of a fluorescence-linked immunosorbent assay (FLISA) based FIS.
- Assessment of FIS performance using control samples, repeatability, and intermediate precision tests.
- Determination of the lower limit of quantification and linearity.
- Comparative analysis of canine plasma CRP levels measured by FIS and an automated analyzer.
Main Results:
- The FIS demonstrated high precision with coefficients of variation below 2.4% (repeatability) and 4.4% (intermediate precision).
- The system achieved a lower limit of quantification of 3.9 mg/L with linearity confirmed up to 465 mg/L.
- A high correlation (R=0.9809) was observed between FIS and automated analyzer measurements, with 38 of 39 samples within acceptable error limits.
Conclusions:
- The developed FIS exhibits analytical accuracy and quantitative capability for canine CRP equivalent to conventional automated systems.
- FIS presents a viable alternative for accurate and reliable C-reactive protein quantification in veterinary diagnostics.
- This technology holds promise for improving diagnostic efficiency in veterinary clinical settings.
Abstract:
This study aimed to construct a measurement system with the same performance as a measurement system using an automated analyzer and immunoturbidimetric reagents (comparative method) using a flow-type immunosensor (FIS) based on the fluorescence-linked immunosorbent assay technology. In the FIS constructed in this study, all control samples were within the indicated values. The coefficient of variation of repeatability and intermediate precision were less than 2.4% and less than 4.4%, respectively. The lower limit of quantification in this measurement system was 3.9 mg/L, and linearity was confirmed for quantification values, ranging from 3.9 to 465 mg/L. Canine plasma samples (N = 39) were used to measure C-reactive protein (CRP) levels using the comparative method (x) and FIS (y). The regression equation between the measurements was y = 1.035 × - 0.002, with a correlation coefficient of 0.9809, indicating a significantly high correlation. Although the Brandt-Altman analysis suggested the possibility of a proportional systematic error between the two measurements, 38 of the 39 canine plasma samples measured fell within the acceptable range of error, indicating that the measurements are highly consistent. These results suggest that the analytical accuracy of the FIS constructed in this study and the quantitative value of canine CRP are equivalent to those of measurement systems using automated analyzers and immunoturbidimetric reagents.
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