分析化学2011，Vol.39Issue(3)：300-304,5.DOI:10.3724/SP.J.1096.2011.00300
核酸适配体识别-荧光法检测赭曲霉素A
An Aptamer-based Fluorescence Assay for Ochratoxin A
摘要
Abstract
A novel analytical method for the detection of Ochratoxin A(OTA) was established based on the aptamer recognition and fluorescent probe technology. The method was developed according to the fact that when the immobilized aptamer bond to the target OTA, it can induce the conformation change of aptamer and result in the dissociation of the carboxy-fluorescein(FAM)-tagged complementary DNA chain from aptamer, and finally lead to the fluorescent signals change. Based on it, OTA can be quantified. All the condition factors affecting the performance of the present method were investigated. The results show if the avidin concentration coated on the microplate is 25 mmg/L, the aptamer concentration is 50 nmol/L, the concentration of FAM-tagged complementary DNA chain is 150 nmol/L, 10 mmol/L n-2-hydroxyethyl-piperazine-n-ethane sulfonic acid(HEPES) pH 7.0(contain 120 mmol/L NaCl, 5 mmol/L KCl, 20 mmol/L MgC12, 20 mmol/L CaCl2) was chosen as the binding buffer solution and the binding reaction was conducted at 45 ℃ for 40 min, the optimal analytical performance can be achieved. Under the optimal conditions, the linear range for the OTA concentration detection is 2.0 × 10- 11-1.0 × 10-8 g/mL with a detection limit of 1 × 10-11 g/mL. The RSD is 2.6％ for 11 parallel measurements of 1 × 10-9 g/mL OTA. Meanwhile, the present method is highly selective for OTA and easy to be operated. It has been successfully applied to measure OTA content in real samples.关键词
核酸适配体/荧光/分子识别/赭曲霉素AKey words
Aptamer/ Fluorescence analysis/ Molecular recognition/ Ochratoxin A引用本文复制引用
段诺,吴世嘉,王周平..核酸适配体识别-荧光法检测赭曲霉素A[J].分析化学,2011,39(3):300-304,5.基金项目
本文系国家"863"计划(No.2008AA10Z419)、国家自然科学基金(No.20805019)、教育部博士点基金新教师计划(No.20070295014)、江苏省自然科学基金(No.BK20081603)、"111"引智计划((No.B07029)资助项目 （No.2008AA10Z419）
