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Self-assembly and label-free fluorescent aptasensor based on deoxyribonucleic acid intercalated dyes for detecting lactoferrin in milk powder

文献类型： 外文期刊

作者： Liu, Jiahui 1 ; Li, Tengfei 3 ; Qin, Hongwei 1 ; Li, Linsen 4 ; Yan, Mengmeng 1 ; Zhu, Chao 1 ; Qu, Feng 4 ; Abd El-Aty, A. M. 5 ;

作者机构： 1.Shandong Acad Agr Sci, Inst Qual Stand & Testing Technol Agro Prod, Jinan, Peoples R China

2.Shandong Prov Key Lab Test Technol Food Qual & Saf, Jinan, Peoples R China

3.Hebei Univ Engn, Coll Life Sci & Food Engn, Handan, Peoples R China

4.Beijing Inst Technol, Sch Life Sci, Key Lab Mol Med & Biotherapy, Beijing, Peoples R China

5.Qilu Univ Technol, Shandong Acad Sci, State Key Lab Biobased Mat & Green Papermaking, Jinan, Peoples R China

6.Cairo Univ, Fac Vet Med, Dept Pharmacol, Giza, Egypt

7.Ataturk Univ, Med Fac, Dept Med Pharmacol, Erzurum, Turkey

关键词： aptamer; fluorescent aptasensor; DNA intercalated dyes; lactoferrin detection; milk powder

期刊名称：FRONTIERS IN NUTRITION （ 影响因子：6.59； 五年影响因子：6.873 ）

ISSN： 2296-861X

年卷期： 2022 年 9 卷

页码：

收录情况： SCI

摘要： Lactoferrin (Lf), an iron-binding glycoprotein, regulates the immune system. It has broad-spectrum antimicrobial activity and is critical for child physical growth and development. As a common additive in the dairy industry, it is crucial to quantify LF content. This study established a self-assembly and universal fluorescence aptasensor for detecting LF in milk powder based on structure-selective dyes of PicoGreen intercalated in the label-free aptamer. Herein, the aptamer functions as both a specific recognition element against targets and a fluorescent signal reporter integrated with structure-selective dyes. First, the aptamer folds into a three-dimensional spatial structure based on complementary base pairings and intermolecular weak non-covalent interactions. Then, the dye is intercalated into the minor groove structures of the aptamer and triggers its potential fluorescent property. When the target exists, the aptamer binds to it preferentially, and its space structure unfolds. This causes the freeing of the subsequent dye and decreases the corresponding fluorescence. Hence, the reflected fluorescence signals could directly determine the target concentrations. Under the optimum conditions, a good linear relationship (R-2, 0.980) was obtained in the Lf range from 20 to 500 nM with a detection limit of 3 nM (2.4 mg/kg) and good specificity, as well as a reliable recovery of 95.8-105.1% in milk powder. In addition, the universality was also confirmed with a good performance by quickly changing the aptamers against other targets (chlorpyrifos, acetamiprid, bovine thyroglobulin, and human transferrin) or utilizing another fluorescence dye. Therefore, this self-assembly aptasensor provides a universal and concise strategy for effective detection.

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