期刊
THERANOSTICS
卷 8, 期 12, 页码 3189-3197出版社
IVYSPRING INT PUBL
DOI: 10.7150/thno.24034
关键词
point-of-care testing; immunochromatographic assay; hook effect; C-reactive protein; wide range detection; minute sample volume
资金
- Mid-career Researcher Program through National Research Foundation grant - Ministry of Science, ICT and Future Planning [NRF-2017R1A2B3010816]
- R&D Program for Society of the National Research Foundation (NRF) - Ministry of Science, ICT & Future Planning [2015M3A9E2031375]
- Original Technology Research Program for Brain Science through the National Research Foundation of Korea (NRF) [NRF-2015M3C7A1029196]
The immunochromatographic (ICA) assay is a highly promising platform for rapid and simple detection of C-reactive protein (CRP) which is an indicator of the different phases of various diseases, as well as of inflammation and infection. However, the hook effect in the ICA assay limits the quantification of CRP levels at high CRP concentrations. Methods: In this study, we developed a hook effect-free immunochromatographic assay (HEF-ICA) to detect CRP over a wide concentration range. The hook effect results from the simultaneous reaction of an excess target antigens with both immobilized and labeled antibodies respectively. To reduce the potential occurrence of this simultaneous reaction, we separated the migration of the target antigen and gold nanoparticle (AuNP)-labeled antibodies on a nitrocellulose membrane and analyzed the time profiles by modifying the ICA structure. Results: The signal intensity of HEF-ICA was saturated at high CRP concentrations, without decreasing. The titration curve of HEF-ICA was adjusted with the Hill equation, and HEF-ICA was performed with the following parameters: limit of detection, 43 ng mL(-1); dynamic range, 119 ng mL(-1) to 100 mu g mL(-1). The accuracy of the newly developed assay was evaluated using 33 clinical samples via comparison with a clinical chemistry analyzer. Conclusion: HEF-ICA enabled the measurement of a wide range of CRP concentrations without the hook effect, and was suitable for point-of-care testing with fingertip blood sampling, as only a minute sample volume (2.5 mu L) was required.
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