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28. A Metal Organic Framework-Based Light Scattering ELISA for the Detection of Staphylococcal Enterotoxin B.

Author

Mao, Kai, Tian, Lili, Luo, Yujie, Li, Qian, Chen, Xi, Zhan, Lei, Li, Yuanfang, Huang, Chengzhi, and Zhen, Shujun

Subjects
LIGHT scattering, ORGANOMETALLIC compounds, ENTEROTOXINS, ENZYME-linked immunosorbent assay, FOOD poisoning, POROSITY
Abstract

Enzyme-linked immunosorbent assay (ELISA) is one of the most commonly used method for the detection of staphylococcal enterotoxin B (SEB), the main protein toxin causing staphylococcal food poisoning. However, the traditional ELISA reaction needs to be stopped by sulfuric acid to obtain stable colorimetric signal, and it is easily influenced by a colored sample. In order to address this problem, a new ELISA method using zeolite imidazolate skeleton-8 metal-organic framework (ZIF-8 MOF) as a light scattering (LS) reporter for SEB detection was developed in this work. ZIF-8 MOF has the characteristics of high porosity, large specific surface area, clear pore structure, and adjustable size, which is one of the most representative MOFs constructed from Zn2+ and 2-methylimidazole (2-mIM). The 2-mIM ligand of ZIF-8 exhibited antioxidant activity and can strongly react with H2O2, which could destroy the structure of ZIF-8, resulting in the obvious decrease in LS intensity. We combined this specific reaction with the sandwich immune reaction to construct the LS ELISA method for the successful detection of SEB. This method is more reliable than commercial tests kits for the detection of colored samples, and it is simple, sensitive, and selective, and has great potential in the detection of other toxins by simply changing the corresponding recognition units. [ABSTRACT FROM AUTHOR]

Published
2023
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31. Catalyzed Hairpin Assembly-Assisted DNA Dendrimer Enhanced Fluorescence Anisotropy for MicroRNA Detection.

Author

Xie, Tianjin, Liu, Yuxin, Xie, Jiali, Luo, Yujie, Mao, Kai, Huang, Chengzhi, Li, Yuanfang, and Zhen, Shujun

Subjects
FLUORESCENCE anisotropy, NUCLEIC acid probes, MOLECULAR volume, HAIRPIN (Genetics), DENDRIMERS, POLYAMIDOAMINE dendrimers, DNA
Abstract

Biomacromolecules have been employed successfully as fluorescence anisotropy (FA) amplifiers for biosensing in reported studies. However, the sensitivities of the traditional biomacromolecule amplified FA strategies need to be improved because of the relatively low molecular weight or volume of a single biomacromolecule and the 1:1 binding ratio between the fluorophore-linked probe and target. In this work, a DNA dendrimer with a high molecular weight and volume was employed as a new FA amplifier, which was coupled with target-catalyzed hairpin assembly (CHA) for the sensitive detection of miRNA-21. The fluorophore-modified probe DNA (pDNA) was fixed on the DNA dendrimer, resulting in a high FA value. The addition of miRNA-21 triggered the CHA process and produced plenty of H1-H2 hybrids. The complex of H1-H2 bound to the DNA dendrimer and released the pDNA through a toehold-mediated strand exchange reaction. Thus, a low FA value was obtained because of the low mass and volume of free pDNA. Based on the dramatically reduced FA, miRNA-21 was detected in the range of 1.0–19.0 nM and the limit of detection was 52.0 pM. In addition, our method has been successfully utilized for miRNA-21 detection in human serum. This strategy is sensitive and selective and is expected to be used to detect other biomolecules simply by changing the corresponding nucleic acid probe. [ABSTRACT FROM AUTHOR]

Published
2022
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