Anti-Idiotypic Nanobody-Phage Based Real-Time Immuno-PCR for Detection of Hepatocarcinogen Aflatoxin in Grains and Feedstuffs

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• 作者：Jiawen Lei ; Peiwu Li ; Qi Zhang ; Yanru Wang ; Zhaowei Zhang ; Xiaoxia Ding ; Wen Zang
• 刊名：Analytical Chemistry
• 出版年：2014
• 出版时间：November 4, 2014
• 年：2014
• 卷：86
• 期：21
• 页码：10841-10846
• 全文大小：348K
• ISSN：1520-6882

文摘

Aflatoxins are a group of extremely toxic small molecules that have been involved in human hepatic and extrahepatic carcinogenesis as causative agents. Herein, we developed a real-time immuno polymerase chain reaction (IPCR) assay for the accurately quantitative detection of aflatoxins in agri-products base on a M13 phage containing aflatoxin anti-idiotypic nanobody and its encoding DNA which was used to design the specific primers. The limit of detection (LOD) of the assay is 0.02 ng/mL, which exhibits a 4-fold improvement over traditional phage ELISA. The developed method was successfully validated with the samples of corn, rice, peanut, and feedstuff, which are major aflatoxin-contaminated agri-products. And the recoveries were from 77.05 to 122.16%. For further validation, the developed assay was also compared with a reference HPLC method for the analysis of aflatoxins in corn and peanuts, and concordant results (R^2聽= 0.991) were obtained. In this context, this study provides a novel opportunity to analyze aflatoxins in agri-products.