基于核酸碱基猝灭荧光团的核酸适配体传感器检测赭曲霉毒素A

引用本文: 曲瑶, 张亚旗, 肖光, 杨成. 基于核酸碱基猝灭荧光团的核酸适配体传感器检测赭曲霉毒素A[J]. 分析化学, 2020, 48(10): 1409-1415. doi: 10.19756/j.issn.0253-3820.201260 shu

Citation: QU Yao, ZHANG Ya-Qi, XIAO Guang, YANG Cheng. Aptasensor Based on Nucleic Acid Base Quenching Fluorophore for Detection of Ochratoxin A[J]. Chinese Journal of Analytical Chemistry, 2020, 48(10): 1409-1415. doi: 10.19756/j.issn.0253-3820.201260 shu

基于核酸碱基猝灭荧光团的核酸适配体传感器检测赭曲霉毒素A

大连理工大学化工学院化学系, 精细化工国家重点实验室, 大连 116024
基金项目:
本文系中央高校基本科研业务费专项资金（No.DUT15RC（3）064）、辽宁省博士科研启动基金（No.201501167）和电分析化学国家重点实验室开放课题（No.SKLEAC201604）资助

摘要: 赭曲霉毒素A（OTA）是农产品中常见的霉菌毒素，对人和动物具有较强的毒性，以及致畸、致癌、致突变作用，因此，建立简单、快速、低成本、高灵敏的OTA检测方法是即时农产品质量监测和保障消费者安全的有效手段。本研究以鸟嘌呤碱基为猝灭剂，以单标记荧光素（FAM）的寡聚核酸为探针，基于核酸适配体构建了生物传感器，用于检测葡萄酒中的OTA。鸟嘌呤是具有多个给电子基团的稠杂环化合物，电子密度较大，在核酸碱基中氧化电位最低，最容易被氧化，作为电子供体可通过光诱导电子转移过程猝灭荧光基团。OTA不存在时，标记有FAM的寡聚核酸探针与OTA核酸适配体杂交，FAM靠近鸟嘌呤荧光被猝灭；样品中存在OTA时，其与核酸适配体特异性结合，形成四链体结构，抑制寡聚核酸探针与适配体结合，FAM的荧光得以恢复，通过FAM荧光强度的恢复率实现对OTA的检测。本方法检测OTA的线性范围为0.67~7.80 nmol/L，检出限为0.67 nmol/L（0.27 μg/kg，S/N=3）。实际红酒样品中OTA的加标回收率为92.4%~100.9%。与纳米金、单壁碳纳米管、氧化石墨烯等纳米材料为猝灭剂的方法相比，本方法具有检测成本低、选择性好、检出限低等优点。

关键词:

English

Aptasensor Based on Nucleic Acid Base Quenching Fluorophore for Detection of Ochratoxin A

State Key Laboratory of Fine Chemicals, School of Chemistry, College of Chemical Engineering, Dalian University of Technology, Dalian 116024, China
Received Date: 27 May 2020
Revised Date: 06 July 2020
Fund Project:
This work was supported by the Fundamental Research Funds for the Central Universities (No. DUT15RC(3) 064),the Scientific Research Foundation for Doctoral Research of Liaoning Province, China (No. 201501167) and the Open Funds of the State Key Laboratory of Electroanalytical Chemistry (No. SKLEAC201604).

Abstract: Ochratoxin A (OTA) is one kind of mycotoxin, which is toxic and widely distributed. To avoid the poison of OTA, it is necessary to develop a rapid, low-cost and highly sensitive detection method to evaluate the concentration of OTA in the food. Here, an aptamer biosensor was developed to detect OTA in wine using single labeled oligonucleotide with fluorescein (FAM) as probe and guanine in the aptamer as quencher. Guanine was a heterocyclic compound that had a high electron density and the lowest oxidation potential in nucleic acid bases. Guanine was easily to be oxidized. As an electron donor, the fluorophore could be quenched by guanine through the process of photoinduced electron transfer (PET). When the probe labeled with FAM hybridized with the OTA aptamer, the fluorescence of FAM was quenched by the guanine in OTA aptamer. In the presence of OTA, due to the specific interaction between OTA and aptamer, OTA induced aptamer folding to G-quadruplexes, which inhibited the binding of probe with aptamer, and thus led to the fluorescence intensity recovery. The concentration of OTA could be determined by the recovery ratio of FAM fluorescence intensity. The linear range for detection of OTA by this aptasenor was 0.67-7.80 nmol/L, the linear calibration equation was R(%)=-0.02+1.76C_OTA (nmol/L), and LOD was 0.67 nmol/L (0.27 g/kg, S/N=3).The recoveries of OTA in red wine sample were in the range of 92.4%-100.9%. Compared with sensing strategies based on gold nanoparticles, single-walled nanotubes and graphene oxide, this method had many advantages such as low cost, good selectivity and high sensitivity.

Key words:

1. [1]
  de Girolamo A, Mckeague M, Miller J D, Derosa M C, Visconti A. Food Chem., 2011, 127(3):1378-1384
2. [2]
  GB 2761-2017, National Food Safety Standard-Maximum Levels of Mycotoxins in Foods, National Standards of the People's Republic of China 食品安全国家标准, 食品中真菌毒素限量. 中华人民共和国国家标准. GB 2761-2017
3. [3]
  Campone L, Piccinelli A L, Celano R, Pagano I, Russo M, Rastrelli L. Food Chem., 2018, 244:128-135
4. [4]
  Sakin F, Tekeli I O, Yipel M, Kurekci C. Food Control, 2018, 90:317-323
5. [5]
  Prieto-Simón B, Campàs M, Marty J, Noguer T. Biosens. Bioelectron., 2008, 23(7):995-1002
6. [6]
  Li P, Zhang Q, Zhang W. TrAC-Trends Anal. Chem., 2009, 28(9):1115-1126
7. [7]
  Wilson D S, Szostak J W. Annu. Rev. Biochem., 1999, 68:611-647
8. [8]
  Liu J, Cao Z, Lu Y. Chem. Rev., 2009, 109(5):1948-1998
9. [9]
  WANG Zi-Jian, CHEN Er-Ning, YANG Ge, ZHAO Xin-Ying, QU Feng. Chinese J. Anal. Chem., 2020, 48(5):573-582 王子健, 陈尔凝, 杨歌, 赵新颖, 屈锋. 分析化学, 2020, 48(5):573-582
10. [10]
  Lin C, Zheng H, Sun M, Guo Y, Luo F, Guo L, Qiu B, Lin Z, Chen G. Anal. Chim. Acta, 2018, 1002:90-96
11. [11]
  Lu Z, Chen X, Hu W. Sens. Actuators B, 2017, 246:61-67
12. [12]
  Kuang H, Chen W, Xu D, Xu L, Zhu Y, Liu L, Chu H, Peng C, Xu C, Zhu S. Biosens. Bioelectron., 2010, 26(2):710-716
13. [13]
  Shen P, Li W, Liu Y, Ding Z, Deng Y, Zhu X, Jin Y, Li Y, Li J, Zheng T. Anal. Chem., 2017, 89(21):11862-11868
14. [14]
  Shen P, Li W, Ding Z, Deng Y, Liu Y, Zhu X, Cai T, Li J, Zheng T. Anal. Biochem., 2018, 554:28-33
15. [15]
  Mckeague M, Velu R, Hill K, Bardóczy V, Mészáros T, Derosa M. Toxins, 2014, 6(8):2435-2452
16. [16]
  Cruz-Aguado J A, Penner G. Anal. Chem., 2008, 80(22):8853-8855
17. [17]
  Geng X, Zhang D, Wang H, Zhao Q. Anal. Bioanal. Chem., 2013, 405(8):2443-2449
18. [18]
  Lv X, Zhang Y, Liu G, Du L, Wang S. RSC Adv., 2017, 7(27):16290-16294
19. [19]
  Guo Z, Ren J, Wang J, Wang E. Talanta, 2011, 85(5):2517-2521
20. [20]
  Sheng L, Ren J, Miao Y, Wang J, Wang E. Biosens. Bioelectron., 2011, 26(8):3494-3499
21. [21]
  Lu Z, Chen X, Hu W. Sens. Actuators B, 2017, 246:61-67
22. [22]
  Torimura M, Kurata S, Yamada K, Yokomaku T, Kamagata Y, Kanagawa T, Kurane R. Anal. Sci., 2001, 17(1):155-160
23. [23]
  Mao H, Luo G, Zhan Y, Zhang J, Yao S, Yu Y. Analyst, 2018, 143(14):3292-3301
24. [24]
  Fan X, Lin F, Zhang Y, Zhao J, Li H, Yao S. New J. Chem., 2012, 36(11):226-2265
25. [25]
  Wang W, Chen C, Qian M X, Zhao X S. Sens. Actuators B, 2008, 129(1):211-217
26. [26]
  Xiang D S, Zhai K, Wang L Z. Analyst, 2013, 138(18):5318-5324
27. [27]
  Yang C, Wang Y, Marty J, Yang X. Biosens. Bioelectron., 2011, 26(5):2724-2727
28. [28]
  Yang C, Lates V, Prieto-Simón B, Marty J, Yang X. Talanta, 2013, 116:520-526
29. [29]
  Cruz-Aguado J A, Penner G. J. Agr. Food Chem., 2008, 56(22):10456-10461

扫一扫看文章

计量

PDF下载量: 11
文章访问数: 1001
HTML全文浏览量: 72

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

基于核酸碱基猝灭荧光团的核酸适配体传感器检测赭曲霉毒素A

English

Aptasensor Based on Nucleic Acid Base Quenching Fluorophore for Detection of Ochratoxin A

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于核酸碱基猝灭荧光团的核酸适配体传感器检测赭曲霉毒素A

English

Aptasensor Based on Nucleic Acid Base Quenching Fluorophore for Detection of Ochratoxin A

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs