Abstract
A fluorescence enhancement method is presented for the determination of ochratoxin A (OTA). The interaction of OTA with its aptamer causes structural changes which, in turn, change fluorescence of enzymatically generated polythymine-coated copper nanoparticles (CuNPs) (with excitation/emission maxima at 340/625 nm). The OTA-binding aptamer was immobilized on magnetic beads. When it binds OTA, it is partially released and exposes a region with a partly complimentary DNA strand (cDNA). After magnetic separation, the cDNA was employed as a primer to trigger the terminal deoxynucleotidyl transferase-mediated polymerization. This process generates polythymine which act as a template for synthesis of the CuNPs. The method is sensitive in having a 2.0 nM detection limit for OTA. It was successfully applied to the determination of OTA in spiked diluted red wine.
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References
Bayman P, Baker JL, Doster MA, Michailides TJ, Mahoney NE (2002) Ochratoxin production by the Aspergillus ochraceus group and Aspergillus alliaceus. Appl Environ Microbiol 68(5):2326–2329
Duarte SC, Pena A, Lino CM (2010) A review on ochratoxin A occurrence and effects of processing of cereal and cereal derived food products. Food Microbiol 27(2):187–198
Petzinger E, Ziegler K (2000) Ochratoxin A from a toxicological perspective. J Vet Pharmacol Ther 23(2):91–98
Bauer J, Gareis M (1987) Ochratoxin-a in the food-chain. J Vet Med 34(8):613–627
Pfohl-Leszkowicz A, Manderville RA (2007) Ochratoxin A: an overview on toxicity and carcinogenicity in animals and humans. Mol Nutr Food Res 51(1):61–99
Liu JW, Cao ZH, Lu Y (2009) Functional nucleic acid sensors. Chem Rev 109(5):1948–1998
Cruz-Aguado JA, Penner G (2008) Determination of Ochratoxin A with a DNA aptamer. J Agric Food Chem 56(22):10456–10461
Wang Y, Ning G, Wu Y, Wu S, Zeng B, Liu G, He X, Wang K (2018) Facile combination of beta-cyclodextrin host-guest recognition with exonuclease-assistant signal amplification for sensitive electrochemical assay of ochratoxin A. Biosens Bioelectron 124-125:82–88
Zhou L, Sun N, Xu L, Chen X, Cheng H, Wang J, Pei R (2016) Dual signal amplification by an “on-command” pure DNA hydrogel encapsulating HRP for colorimetric detection of ochratoxin A. RSC Adv 6(115):114500–114504
Lin C, Zheng H, Sun M, Guo Y, Luo F, Guo L, Qiu B, Lin Z, Chen G (2018) Highly sensitive colorimetric aptasensor for ochratoxin A detection based on enzyme-encapsulated liposome. Anal Chim Acta 1002:90–96
Lee J, Jeon CH, Ahn SJ, Ha TH (2014) Highly stable colorimetric aptamer sensors for detection of ochratoxin A through optimizing the sequence with the covalent conjugation of hemin. Analyst 139(7):1622–1627
Zhou Z, Hao N, Zhang Y, Hua R, Qian J, Liu Q, Li H, Zhu W, Wang K (2017) A novel universal colorimetric sensor for simultaneous dual target detection through DNA-directed self-assembly of graphene oxide and magnetic separation. Chem Commun (Camb) 53(52):7096–7099
Gillibert R, Triba MN, Lamy de la Chapelle M (2017) Surface enhanced Raman scattering sensor for highly sensitive and selective detection of ochratoxin A. Analyst 143(1):339–345
Zhang G, Zhu C, Huang Y, Yan J, Chen A (2018) A lateral flow strip based Aptasensor for detection of Ochratoxin A in corn samples. Molecules 23(2):291
Lu Z, Chen X, Hu W (2017) A fluorescence aptasensor based on semiconductor quantum dots and MoS2 nanosheets for ochratoxin A detection. Sensors Actuators B Chem 246:61–67
Lv L, Li D, Cui C, Zhao Y, Guo Z (2017) Nuclease-aided target recycling signal amplification strategy for ochratoxin A monitoring. Biosens Bioelectron 87:136–141
Lv L, Li D, Liu R, Cui C, Guo Z (2017) Label-free aptasensor for ochratoxin A detection using SYBR gold as a probe. Sensors Actuators B Chem 246:647–652
Song C, Hong W, Zhang X, Lu Y (2018) Label-free and sensitive detection of Ochratoxin A based on dsDNA-templated copper nanoparticles and exonuclease-catalyzed target recycling amplification. Analyst 143:1829–1834
Dai S, Wu S, Duan N, Wang Z (2016) A luminescence resonance energy transfer based aptasensor for the mycotoxin Ochratoxin A using upconversion nanoparticles and gold nanorods. Microchim Acta 183(6):1909–1916
Liu Y, Yan H, Shangguan J, Yang X, Wang M, Liu W (2018) A fluorometric aptamer-based assay for ochratoxin A using magnetic separation and a cationic conjugated fluorescent polymer. Microchim Acta 185(9):254
Rotaru A, Dutta S, Jentzsch E, Gothelf K, Mokhir A (2010) Selective dsDNA-templated formation of copper nanoparticles in solution. Angew Chem Int 49(33):5665–5667
Song C, Yang X, Wang K, Wang Q, Huang J, Liu J, Liu W, Liu P (2014) Label-free and non-enzymatic detection of DNA based on hybridization chain reaction amplification and dsDNA-templated copper nanoparticles. Anal Chim Acta 827:74–79
Zhang LL, Zhao JJ, Duan M, Zhang H, Jiang JH, Yu RQ (2013) Inhibition of dsDNA-templated copper nanoparticles by pyrophosphate as a label-free fluorescent strategy for alkaline phosphatase assay. Anal Chem 85(8):3797–3801
Chen JH, Liu J, Fang ZY, Zeng LW (2012) Random dsDNA-templated formation of copper nanoparticles as novel fluorescence probes for label-free lead ions detection. Chem Commun 48:1057–1059
Liu H, Ma C, Wang J, Wang K, Wu K (2017) A turn-on fluorescent method for determination of the activity of alkaline phosphatase based on dsDNA-templated copper nanoparticles and exonuclease based amplification. Microchim Acta 184(7):2483–2488
Qing ZH, He XX, He DG, Wang KM, Xu FZ, Qing TP, Yang X (2013) Poly(thymine)-templated selective formation of fluorescent copper nanoparticles. Angew Chem Int 52(37):9719–9722
Ye T, Li C, Su C, Ji X, Zheng J, Tinnefeld P, He Z (2015) Enzymatic polymerization of poly(thymine) for the synthesis of copper nanoparticles with tunable size and their application in enzyme sensing. Chem Commun (Camb) 51(41):8644–8647
Luo L, Xu F, Shi H, He X, Qing T, Lei Y, Tang J, He D, Wang K (2017) Label-free and sensitive assay for deoxyribonuclease I activity based on enzymatically-polymerized superlong poly(thymine)-hosted fluorescent copper nanoparticles. Talanta 169:57–63
Chen J, Xu Y, Ji X, He Z (2017) Enzymatic polymerization-based formation of fluorescent copper nanoparticles for the nuclease assay. Sensors Actuators B Chem 239:262–269
Cao J, Wang W, Bo B, Mao X, Wang K, Zhu X (2017) A dual-signal strategy for the solid detection of both small molecules and proteins based on magnetic separation and highly fluorescent copper nanoclusters. Biosens Bioelectron 90:534–541
Hu W, Ning Y, Kong J, Zhang X (2015) Formation of copper nanoparticles on poly(thymine) through surface-initiated enzymatic polymerization and its application for DNA detection. Analyst 140(16):5678–5684
Covarelli L, Beccari G, Marini A, Tosi L (2012) A review on the occurrence and control of ochratoxigenic fungal species and ochratoxin A in dehydrated grapes, non-fortified dessert wines and dried vine fruit in the Mediterranean area. Food Control 26(2):347–356
Acknowledgements
The authors thank the Natural Science Foundation Project of CQ (No. cstc2018jscx-msybX0263), National Natural Science Foundation of China (No. 21405125), China Agriculture Research System (No. CARS-27) and National Risk Assessment Program for Agricultural Products Quality and Safety (No. GJFP2018003 and GJFP2017013) for financial support.
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He, Y., Tian, F., Zhou, J. et al. A fluorescent aptasensor for ochratoxin A detection based on enzymatically generated copper nanoparticles with a polythymine scaffold. Microchim Acta 186, 199 (2019). https://doi.org/10.1007/s00604-019-3314-z
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DOI: https://doi.org/10.1007/s00604-019-3314-z