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一种基于特异性适配体和杂交链式反应生物放大作用检测黄曲霉毒素B1的电化学生物传感器。

An electrochemical biosensor for the detection of aflatoxin B1 based on the specific aptamer and HCR biological magnification.

作者信息

Zhang Hongyan, Ye Siying, Huang Lishan, Fan Shen, Mao Weiwei, Hu Yijin, Yu Yuyan, Fu Fengfu

机构信息

Joint National Local Engineering Research Center of Fujian and Taiwan Chinese Medicine Molecular Biotechnology, College of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350122, China.

Key Lab of Analysis and Detection for Food Safety of Ministry of Education, Fujian Provincial Key Lab of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China.

出版信息

Anal Methods. 2022 Dec 22;15(1):99-108. doi: 10.1039/d2ay01682f.

Abstract

Aflatoxin B1 (AFB1) is a highly toxic mycotoxin, which causes severe acute or cumulative poisoning. Therefore, it is important to develop sensitive and selective detection methods for AFB1 for the safety of food and medicinal herbs. Herein, we have developed a "signal-on" electrochemical aptasensor based on the high specificity of the aptamer and hybridization chain reaction (HCR) biological amplification for AFB1 detection. In this work, thiol-modified complementary DNA (cDNA) immobilized on the surface of a gold electrode (GE) served as an initiator DNA. When AFB1 was present, it competed with the cDNA for binding to the aptamers, which resulted in the detaching of aptamers from the cDNA-aptamer duplexes. Then, the single-stranded cDNA acted as an initiator to trigger the HCR signal amplification. Therefore, long double-stranded DNA (dsDNA) products were produced, which could load large amounts of methylene blue (MB) molecules to generate a distinct electrochemical signal. Under the optimized conditions, the proposed electrochemical aptasensor achieved the ultrasensitive detection of AFB1 with a linear detection range of 0.01-100 pg mL, and a limit of detection (LOD) down to 2.84 fg mL. Furthermore, the electrochemical aptasensor was successfully applied for detecting AFB1 in corn and two kinds of traditional Chinese medicine samples, indicating the potential value for AFB1 detection in practical samples.

摘要

黄曲霉毒素B1(AFB1)是一种剧毒霉菌毒素,可导致严重的急性或累积性中毒。因此,为了食品和草药的安全,开发灵敏且具选择性的AFB1检测方法很重要。在此,我们基于适体的高特异性和杂交链式反应(HCR)生物扩增技术,开发了一种用于AFB1检测的“信号开启”型电化学适体传感器。在这项工作中,固定在金电极(GE)表面的硫醇修饰互补DNA(cDNA)用作引发DNA。当存在AFB1时,它与cDNA竞争结合适体,导致适体从cDNA - 适体双链体上脱离。然后,单链cDNA作为引发剂触发HCR信号放大。因此,产生了长双链DNA(dsDNA)产物,其可以负载大量亚甲基蓝(MB)分子以产生明显的电化学信号。在优化条件下,所提出的电化学适体传感器实现了对AFB1的超灵敏检测,线性检测范围为0.01 - 100 pg/mL,检测限(LOD)低至2.84 fg/mL。此外,该电化学适体传感器成功应用于玉米和两种中药样品中AFB1的检测,表明其在实际样品中检测AFB1的潜在价值。

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