Maeno, Hinata and Wong, Pooi-Fong and AbuBakar, Sazaly and Yang, Ming and Sam, Sing-Sin and Jamil-Abd, Juraina and Shunmugarajoo, Anusha and Mustafa, Mahiran and Said, Rosaida Md and Mageswaren, Eashwary and Azmel, Azureen and Mat Jelani, Anilawati (2021) A 3D microfluidic elisa for the detection of severe dengue: sensitivity improvement and vroman effect amelioration by edc-nhs surface modification. Micromachines, 12 (12). ISSN 2072-666X, DOI https://doi.org/10.3390/mi12121503.
Full text not available from this repository.Abstract
Serum is commonly used as a specimen in immunoassays but the presence of heterophilic antibodies can potentially interfere with the test results. Previously, we have developed a microfluidic device called: 3D Stack for enzyme-linked immunosorbent assay (ELISA). However, its evaluation was limited to detection from a single protein solution. Here, we investigated the sensitivity of the 3D Stack in detecting a severe dengue biomarker-soluble CD163 (sCD163)-within the serum matrix. To determine potential interactions with serum matrix, a spike-and-recovery assay was performed, using 3D Stacks with and without surface modification by an EDC-NHS (N-ethyl-N `-(3-(dimethylamino)propyl)carbodiimide/N-hydroxysuccinimide) coupling. Without surface modification, a reduced analyte recovery in proportion to serum concentration was observed because of the Vroman effect, which resulted in competitive displacement of coated capture antibodies by serum proteins with stronger binding affinities. However, EDC-NHS coupling prevented antibody desorption and improved the sensitivity. Subsequent comparison of sCD163 detection using a 3D Stack with EDC-NHS coupling and conventional ELISA in dengue patients' sera revealed a high correlation (R = 0.9298, p < 0.0001) between the two detection platforms. Bland-Altman analysis further revealed insignificant systematic error between the mean differences of the two methods. These data suggest the potentials of the 3D Stack for further development as a detection platform.
| Item Type: | Article |
|---|---|
| Funders: | Ministry of Energy, Science, Technology, Environment and Climate Change (MESTECC), Malaysia [FP0514B0025-2], WorkPackage 5C [GA019-2016], JST A-STEP, Japan [JPMJTM20MD] |
| Uncontrolled Keywords: | ELISA; microfluidics; EDC-NHS coupling |
| Subjects: | Q Science > Q Science (General) Q Science > QD Chemistry |
| Divisions: | Deputy Vice Chancellor (Research & Innovation) Office > Tropical Infectious Diseases Research and Education Centre |
| Depositing User: | Ms Zaharah Ramly |
| Date Deposited: | 01 Jun 2022 02:59 |
| Last Modified: | 01 Jun 2022 02:59 |
| URI: | http://eprints.um.edu.my/id/eprint/34535 |
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