一种基于单克隆抗体的酶联免疫吸附测定法，用于检测稻曲球和水稻样本中的稻曲毒素A。

A monoclonal antibody-based enzyme-linked immunosorbent assay for detection of ustiloxin A in rice false smut balls and rice samples.

作者信息

Fu Xiaoxiang, Wang Xiaohan, Cui Yongliang, Wang Ali, Lai Daowan, Liu Yang, Li Qing X, Wang Baomin, Zhou Ligang

机构信息

College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.

Institute of Agro-products Processing Science and Technology, Chinese Academy of Agricultural Sciences, Beijing 100193, China.

出版信息

Food Chem. 2015 Aug 15;181:140-5. doi: 10.1016/j.foodchem.2015.02.068. Epub 2015 Feb 21.

DOI:10.1016/j.foodchem.2015.02.068

PMID:25794732

Abstract

Ustiloxin A, a cyclopeptide mycotoxin, was isolated from the pathogenic fungus Villosiclava virens that causes rice false smut, a worldwide devastating rice disease. A monoclonal antibody (mAb) 2D3G5 was generated with ustiloxin A-bovine serum albumin conjugate. A highly sensitive and specific indirect competitive enzyme-linked immunosorbent assay (icELISA) was then developed. It possessed a median inhibition concentration (IC50) of 13.8 ng/mL and a working range of 2.8-72 ng/mL. The mAb 2D3G5 recognized ustiloxin B with the cross-reactivity as 4%. The average recoveries of ustiloxin A from rice false smut balls and peeled rice samples ranged from 92% to 117% and from 92% to 107%, respectively. Comparison of the concentrations of ustiloxin A in rice false smut balls detected by both icELISA and high performance liquid chromatography-photodiode array detection indicated that the developed icELISA was suitable for detection of ustiloxin A in rice food and feed samples.

摘要

稻曲毒素A是一种环肽类真菌毒素，从引起稻曲病的病原菌稻绿核菌中分离得到，稻曲病是一种在全球范围内极具破坏性的水稻病害。用稻曲毒素A-牛血清白蛋白偶联物制备了单克隆抗体（mAb）2D3G5。随后建立了一种高灵敏度和特异性的间接竞争酶联免疫吸附测定法（icELISA）。其半数抑制浓度（IC50）为13.8 ng/mL，工作范围为2.8 - 72 ng/mL。单克隆抗体2D3G5对稻曲毒素B有识别能力，交叉反应率为4%。稻曲病菌核和糙米样品中稻曲毒素A的平均回收率分别为92%至117%和92%至107%。通过icELISA和高效液相色谱 - 光电二极管阵列检测法对稻曲病菌核中稻曲毒素A浓度的比较表明，所建立的icELISA适用于检测大米食品和饲料样品中的稻曲毒素A。