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基于杂交链式反应检测赭曲霉毒素A的高效电化学适体传感器研究

发布时间:2018-10-15 14:51
【摘要】:赭曲霉毒素(Ochratoxins,OTs)是一种由赭曲霉、碳黑曲霉和纯绿青霉产生的食源性毒素,主要污染粮谷类农产品,与人类和动物健康关系密切。其中以赭曲霉毒素A(Ochratoxin A,OTA)的毒性最大、散布最广、污染程度最高。本研究利用电化学适体传感器基于杂交链式反应研发设计了一种拥有超低检测限的OTA检测方法。首先,将一段捕获探针固定在金电极上,然后加入与之互补的OTA适配体序列,使其与捕获探针形成DNA杂交链。在环境中存在OTA的情况下,杂交链上的OTA适配体会与游离的OTA结合,脱离电极。然后,加入检测探针和两段生物素标记的互补DNA发夹结构,与电极上剩余的捕获探针结合,并引发杂交链式反应。最后,在电极上滴加碱性磷酸酶(ST-AP),它与DNA链上的生物素结合,通过催化反应底物中的-萘酚磷酸盐(-NP)产生电化学信号,DPV曲线随OTA浓度的增高而增高。在优化条件下,该检测平台对OTA的检测线性范围是0.005-100ng/m L,检测限可达2 pg/m L。此外,该方法在检测谷物样品的OTA应用中,也获得了较为满意的回收率,检测结果与高效液相色谱法(HPLC)相比具有高度的一致性。
[Abstract]:Ochratoxin (Ochratoxins,OTs) is a foodborne toxin produced by Aspergillus ochratus, Aspergillus Niger and Pure Penicillium. It mainly pollutes cereals and is closely related to human and animal health. OTA of ochratoxin A (Ochratoxin was the most toxic, most widespread and the most polluted. In this study, an ultra-low detection limit (OTA) detection method based on hybrid chain reaction was developed using electrochemical aptamer sensor. Firstly, a capture probe was immobilized on the gold electrode, and then the complementary OTA aptamer sequence was added to form the DNA hybridization chain with the capture probe. In the presence of OTA in the environment, the adaptation of OTA on the hybrid chain was associated with the free OTA, and the electrode was detached. Then, the complementary DNA hairpin structure was added with the detection probe and biotinylated probe to combine with the remaining trapping probe on the electrode, and the hybridization chain reaction was initiated. Finally, alkaline phosphatase (ST-AP) was added to the electrode, which binds to the biotin in the DNA chain and produces electrochemical signals by catalyzing-naphthol phosphate (- NP) in the substrate. The DPV curve increases with the increase of OTA concentration. Under the optimized conditions, the linear range of the detection platform for OTA is 0.005-100ng/m L, and the detection limit can reach 2 pg/m L. In addition, the method has been applied to the determination of grain samples with OTA, and the results are in good agreement with that of (HPLC) by high performance liquid chromatography (HPLC).
【学位授予单位】:重庆医科大学
【学位级别】:硕士
【学位授予年份】:2017
【分类号】:TP212.3;R440

【参考文献】

相关期刊论文 前5条

1 张松柏;胡霞;沈广宇;陈瑶;孙琴利;阳明辉;沈国励;俞汝勤;;电化学适体传感器研究进展[J];化学传感器;2014年03期

2 赵丽红;马秋刚;李笑樱;范_g;雷元培;计成;贾亚雄;谢实勇;;抽样调查北京地区猪场饲料及饲料原料赭曲霉毒素A污染状况[J];动物营养学报;2012年10期

3 赖卫华;许杨;熊勇华;陈媛;刘文娟;;赭曲霉毒素A无毒体系胶体金试纸条的研制及与传统胶体金试纸条的比较[J];食品科学;2008年09期

4 赖卫华,熊勇华,陈高明,许杨;应用胶体金试纸条快速检测赭曲霉毒素A的研究[J];食品科学;2005年05期

5 陈雪岚,许杨,袁勇芳;赭曲霉毒素A的酶联免疫吸附法检测[J];卫生研究;2000年06期

相关硕士学位论文 前1条

1 谈敦芳;我国6省区粮谷类食品中赭曲霉毒素A污染水平调查与我国居民膳食暴露评估的研究[D];河北医科大学;2006年



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