Abstract
An accurate and rapid microflow cytometry-based agglutination immunoassay (MCIA) suitable for on-site antibody or antigen detection was proposed. In this study, quantitative C-reactive protein (CRP) detection was chosen as a model assay in order to demonstrate the detection principle. The average transit time was employed to estimate the extent of the agglutination reaction and improve the detection accuracy as compared to the intensity-dependent methods. The detection time was less than 8 min. and only a 20 µL serum sample was needed for each test. The results showed a linear relationship between the average transit time of aggregates and CRP concentrations ranging from 0 to 1 µg/mL. The R2 of this relationship was 0.99. The detection limit of this technology was 0.12 µg/mL CRP. The system used for CRP detection can be extended to also monitor other clinically relevant molecules.
Highlights
Suitable for on-site antibody or antigen detection was proposed
We report a transit microflow cytometry-based agglutination immunoassay (MCIA) for on-site and quantitative detection of antibodies or antigens in serum samples
The experimental setup of the microflow cytometry for C-reactive protein (CRP) detection is shown in Figure 1, and more details of the fabrication process can be found in a previous publication by Benjamin et al [16]
Summary
Suitable for on-site antibody or antigen detection was proposed. In this study, quantitative C-reactive protein (CRP) detection was chosen as a model assay in order to demonstrate the detection principle. The average transit time was employed to estimate the extent of the agglutination reaction and improve the detection accuracy as compared to the intensity-dependent methods. The results showed a linear relationship between the average transit time of aggregates and CRP concentrations ranging from 0 to 1 μg/mL. Particle counting immunoassay is a sensitive and practical tool for the detection of antibodies and antigens. The combination of particle counting assays and microflow cytometry makes on-site immunoassay detection possible. Microflow cytometry was first used for the measurement of aggregates that formed in a particle-based immunoassay in 2003. The intensity of scattering light was employed in order to measure the aggregates. The forward scattering light in microflow cytometry can be applied to distinguish monomers from aggregates in a particle-enhanced immunoassay. Biotinylated bovine serum was detected in order to verify this method with a detection limit of 1.5 pM [4]
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