Development of ZnCdSe/ZnS quantum dot-based fluorescence immunochromatographic assay for the rapid visual and quantitative detection 25⁃hydroxyvitamins D in human serum

Jianfa Wang; Guoshao Sun; Fang Li; Zhi Zhu; Lei Sun; Pengju Lv; Han Yue

doi:10.3389/fbioe.2023.1326254

Development of ZnCdSe/ZnS quantum dot-based fluorescence immunochromatographic assay for the rapid visual and quantitative detection 25⁃hydroxyvitamins D in human serum

Front Bioeng Biotechnol. 2023 Dec 22:11:1326254. doi: 10.3389/fbioe.2023.1326254. eCollection 2023.

Authors

Jianfa Wang¹, Guoshao Sun¹, Fang Li¹, Zhi Zhu¹, Lei Sun², Pengju Lv², Han Yue²

Affiliations

¹ Department of Orthopedics, Zhengzhou Central Hospital Affiliated Zhengzhou University, Zhengzhou, Henan, China.
² Center of Stem Cell and Regenerative Medicine, Zhengzhou Central Hospital Affiliated Zhengzhou University, Zhengzhou, Henan, China.

Abstract

Vitamin D deficiency is associated with various diseases such as obesity, digestive problems, osteoporosis, depression, and infections, and has therefore emerged as a topic of great interest in public healthcare. The quantitative assessment of 25-hydroxyvitamin D (25-OH VD) in human serum may accurately reflect the nutritional status of vitamin D in the human body, which is significant for the prevention and treatment of vitamin D-deficient patients. In this study, we developed an assay for quantitative detection of 25-OH VD based on the 25-OH VD monoclonal antibody (mAb), and identified the optimal process parameters. The following process settings were found to be suitable for the test strips: pH of 7.6, 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) ratio of 1:2000, and the anti-25-OH VD mAb ratio was 1:8. The equilibration time of the immune dynamic assay was 15 min. Under optimal conditions, the quantum dot nanoparticle-based fluorescent immunochromatographic assay (QDs-FICA) exhibited dynamic linear detection of 25-OH VD in PBS, from 5 ng/mL to 100 ng/mL, and the strip quantitative curve could be represented by the following regression equation: y = -0.02088 logx)+1.444 (R2 = 0.9050). The IC50 of the QDs-FICA was 39.6 ± 1.33 ng/mL. The specificity of the QDs-FICA was evaluated by running several structurally related analogues, including 25-OH VD₂, 25-OH VD₃, 1,25-OH₂VD₃, 1,25-OH₂VD₂, VD₂, and VD₃. The coefficients of variation were all below 10%. The shelf life of the test strips in this study was about 160 days at room temperature. Briefly, this study is the first to perform QDs-FICA for the rapid visual and quantitative detection of 25-OH VD, with great potential significance for clinical diagnosis of vitamin D-associated diseases.

Keywords: 25-hydroxyvitamin D; ZnCdSe/ZnS; fluorescence immunochromatographic assay; quantitative detection; quantum dots.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by the henan province medical science and technology project (grant numbers LHGJ20200767), this work was also supported by the Henan Province science and technology research and development projects (grant numbers 222102310032).