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基于纳米颗粒的生物条码技术用于中草药中黄曲霉毒素 B1 的痕量分析。

Nanoparticle based bio-bar code technology for trace analysis of aflatoxin B1 in Chinese herbs.

机构信息

College of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China.

出版信息

J Food Drug Anal. 2018 Apr;26(2):815-822. doi: 10.1016/j.jfda.2017.11.003. Epub 2017 Dec 6.

Abstract

A novel and sensitive assay for aflatoxin B1 (AFB1) detection has been developed by using bio-bar code assay (BCA). The method that relies on polyclonal antibodies encoded with DNA modified gold nanoparticle (NP) and monoclonal antibodies modified magnetic microparticle (MMP), and subsequent detection of amplified target in the form of bio-bar code using a fluorescent quantitative polymerase chain reaction (FQ-PCR) detection method. First, NP probes encoded with DNA that was unique to AFB1, MMP probes with monoclonal antibodies that bind AFB1 specifically were prepared. Then, the MMP-AFB1-NP sandwich compounds were acquired, dehybridization of the oligonucleotides on the nanoparticle surface allows the determination of the presence of AFB1 by identifying the oligonucleotide sequence released from the NP through FQ-PCR detection. The bio-bar code techniques system for detecting AFB1 was established, and the sensitivity limit was about 10 ng/mL, comparable ELISA assays for detecting the same target, it showed that we can detect AFB1 at low attomolar levels with the bio-bar-code amplification approach. This is also the first demonstration of a bio-bar code type assay for the detection of AFB1 in Chinese herbs.

摘要

已开发出一种新颖且灵敏的黄曲霉毒素 B1(AFB1)检测方法,即生物条码分析(BCA)。该方法依赖于经 DNA 编码的多克隆抗体修饰的金纳米颗粒(NP)和经单克隆抗体修饰的磁性微粒(MMP),并随后使用荧光定量聚合酶链反应(FQ-PCR)检测方法检测生物条码形式的放大靶标。首先,制备了独特编码 AFB1 的 DNA 修饰 NP 探针和与 AFB1 特异性结合的单克隆抗体修饰 MMP 探针。然后,获得 MMP-AFB1-NP 三明治复合物,通过 FQ-PCR 检测从 NP 表面解杂交的寡核苷酸来识别从 NP 释放的寡核苷酸序列,从而确定 AFB1 的存在。建立了用于检测 AFB1 的生物条码技术系统,其灵敏度下限约为 10ng/mL,与检测同一目标的 ELISA 检测相当,表明我们可以通过生物条码扩增方法检测到低至飞摩尔级别的 AFB1。这也是首次在中草药中检测 AFB1 的生物条码型检测方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b2e/9322209/30e9411ae791/jfda-26-02-815f1.jpg

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