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使用固定在胺基功能化的树枝状纤维纳米二氧化硅表面的基于适配体的生物传感器测定黄曲霉毒素M1。

Determination of aflatoxin M1 using an aptamer-based biosensor immobilized on the surface of dendritic fibrous nano-silica functionalized by amine groups.

作者信息

Kholafazad Kordasht Houman, Moosavy Mir-Hassan, Hasanzadeh Mohammad, Soleymani Jafar, Mokhtarzadeh Ahad

机构信息

Department of Food Hygiene and Aquatics, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran.

Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

出版信息

Anal Methods. 2019 Aug 1;11(30):3910-3919. doi: 10.1039/c9ay01185d.

Abstract

Aflatoxins are potential food pollutants produced by fungi. Among them, aflatoxin M1 (AF M1) is the most toxic. A great deal of concern is associated with AF M1 toxicity. Aflatoxins are potential food pollutants produced by fungi. Among them, aflatoxin M1 (AF M1) is the most toxic. A great deal of concern is associated with AF M1 toxicity. In the present work, a novel aptamer-based bioassay was developed for monitoring aflatoxin M1 (AF M1) in real samples. A chitosan-modified graphene quantum dot (GQD-CS) nanocomposite was used as a biocompatible substrate coated with dendritic fibrous nanosilica functionalized by amine groups (KCC-1-NH-Tb). Accordingly, an innovative biocompatible polymeric matrix was prepared for aptamer immobilization. The unique oligonucleotide of AF M1 (5'-ATC CGT CAC ACC TGC TCT GAC GCT GGG GTC GAC CCG GAG AAA TGC ATT CCC CTG TGG TGT TGG CTC CCG TAT) labelled by toluidine blue was immobilized on the engineered interface. Hence, a novel aptamer-based bioassay was formed for the highly sensitive quantitation of AF M1 using cyclic voltammetry and differential pulse voltammetry techniques. The structure and morphology of GQDs-CS/KCC-1-NH-Tb was investigated by Fourier transform infrared spectroscopy, X-ray diffraction, atomic force and scanning electron microscopy and energy-dispersive X-ray spectroscopy. The toxicity tests, which were performed by MTT assays, revealed the biocompatible nature of KCC-1-NH-Tb. The engineered aptasensor demonstrated excellent behaviour toward the determination of AF M1, where the low limit of quantification was 10 fM. The proposed aptamer-based bioassay was successfully used for the monitoring of AF M1 in milk samples. This work provides a beneficial reference for the sensing of other toxins in food/pharmaceutical assays and veterinary medicine.

摘要

黄曲霉毒素是由真菌产生的潜在食品污染物。其中,黄曲霉毒素M1(AF M1)毒性最强。AF M1的毒性引发了人们的广泛关注。黄曲霉毒素是由真菌产生的潜在食品污染物。其中,黄曲霉毒素M1(AF M1)毒性最强。AF M1的毒性引发了人们的广泛关注。在本研究中,开发了一种基于新型适配体的生物测定方法,用于监测实际样品中的黄曲霉毒素M1(AF M1)。壳聚糖修饰的石墨烯量子点(GQD-CS)纳米复合材料用作生物相容性基质,表面涂覆有胺基功能化的树枝状纤维纳米二氧化硅(KCC-1-NH-Tb)。据此,制备了一种用于固定适配体的创新型生物相容性聚合物基质。用甲苯胺蓝标记的AF M1独特寡核苷酸(5'-ATC CGT CAC ACC TGC TCT GAC GCT GGG GTC GAC CCG GAG AAA TGC ATT CCC CTG TGG TGT TGG CTC CCG TAT)固定在工程化界面上。因此,形成了一种基于新型适配体的生物测定方法,采用循环伏安法和差分脉冲伏安法技术对AF M1进行高灵敏度定量分析。通过傅里叶变换红外光谱、X射线衍射、原子力显微镜、扫描电子显微镜和能量色散X射线光谱对GQDs-CS/KCC-1-NH-Tb的结构和形貌进行了研究。通过MTT法进行的毒性测试表明KCC-1-NH-Tb具有生物相容性。该工程化适配体传感器在测定AF M1方面表现出色,定量下限为10 fM。所提出的基于适配体的生物测定方法成功用于牛奶样品中AF M1的监测。这项工作为食品/药物分析和兽医学中其他毒素的传感提供了有益的参考。

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