基于适体技术的桔青毒素检测方法的建立

发布时间：2018-03-10 21:54

本文选题：桔青毒素　切入点：指数富集适体系统进化技术　出处：《菌物学报》2016年02期 　论文类型：期刊论文

【摘要】：桔青毒素(citrinin,CTN)是以玉米、谷物、奶酪等为主要成分的食品和动物饲料中常见的、由桔青霉菌产生的酮类真菌毒素,可引起人和动物的慢性中毒或癌症,一直缺少灵敏的快速检测方法。本文通过指数富集适体系统进化技术(简称SELEX)对可能与CTN结合的特异性适体进行了筛选,经过15轮循环,得到17条适体。通过二级结构分析、亲和力检测发现适体13(Apt-13)对CTN有较好的亲和度,解离常数Kd为0.06μmol/L。进一步利用非荧光标记染料Pico Green,利用其与双链DNA结合的原理,建立了桔青毒素非标记荧光检测方法,30min完成检测,最低检测限达到国家标准(50ppb)且与其他毒素无交叉反应。本研究建立的基于适体的桔青毒素检测技术成本低,可以替代传统的基于抗体的检测方法,为霉菌毒素的精准检测技术的开发提供了实验证据。
[Abstract]:Citrinin (CTN) is a common form of food and animal feed made up of corn, grain, cheese, etc. Ketone mycotoxins produced by Penicillium citri can cause chronic poisoning or cancer in humans and animals. There has been a lack of sensitive and rapid detection methods. In this paper, the specific aptamers that may bind to CTN were screened by exponential enrichment aptamer phylogenetic technique (Selex). After 15 cycles, 17 aptamers were obtained. The affinity test showed that aptamer 13apt-13) had good affinity to CTN, and the dissociation constant KD was 0.06 渭 mol / L. further using the principle of non-fluorescent labeling dye Pico Greenand combining with double-stranded DNA, a non-fluorescent detection method for citrinin was established in 30 minutes. The minimum detection limit is up to the national standard of 50 ppb) and there is no cross reaction with other toxins. The technology based on aptamer is low in cost and can replace the traditional antibody based detection method. It provides experimental evidence for the development of precise detection technology of mycotoxins.
【作者单位】：浙江农林大学动物科技学院;浙江大学动物科学学院;
【基金】：国家高技术研究发展计划(863计划)(2012AA101602) 浙江省科学技术厅重大科技专项重大农业项目(2014C02003) 浙江农林大学人才项目(2011FR025;2012FR047;2013FR012;2013FR054)~~
【分类号】：TS207.5;S859.8

【共引文献】