Microchimica Acta

, 185:497 | Cite as

A test strip for ochratoxin A based on the use of aptamer-modified fluorescence upconversion nanoparticles

  • Shijia Wu
  • Lihong Liu
  • Nuo DuanEmail author
  • Wenyue Wang
  • Qianru Yu
  • Zhouping WangEmail author
Original Paper


An aptamer-based test strip is described for visual and instrumental determination of the mycotoxin ochratoxin A (OTA). It is based on the use of NaYF4:Yb,Er upconversion nanoparticles (UCNPs) as a label for the aptamer and on the competition between OTA and its complementary sequence for an OTA-specific aptamer. To improve the analytical performance, the optical properties of the UCNPs, the fluidity of the UCNP-aptamer conjugate, and the migration rate on the nitrocellulose membranes were investigated. Under the optimal experimental conditions and by using a 980-nm laser, the relative fluorescence intensity (test line value/control line value) is proportional to the logarithm of the OTA concentration over a range from 5 to 100 ng·mL−1 (R2 = 0.9955). The limit of the detection is 1.86 ng·mL−1. This aptamer based flow assay can be performed within 15 min and has no serious cross-sensitivity to potentially interfering species. It was successfully applied to the determination of OTA in spiked wheat and beer samples.

Graphical abstract

An aptamer-based upconversion fluorescent strip based on the use of NaYF4:Yb,Er nanoparticles was developed for sensitive detection of Ochratoxin A. The limit of the detection was determined as 1.86 ng·mL−1. The assay can be performed within 15 min, indicating its great potential in point-of-care testing.


Mycotoxin Rapid detection Upconversion nanoparticles Wheat, beer 



This work was partially supported by Key Research and Development Program of Jiangsu Province BE2016306, National Natural Science Fund of China (NSFC 31772086), Project funded by China Postdoctoral Science Foundation (2016 T90430), Project funded by Jiangsu Province Postdoctoral Science Foundation (1601087B, 1701097B), Young Elite Scientists Sponsorship Program by CAST (2017QNRC001) and Fundamental Research Funds for the Central Universities (JUSRP21826).

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2018_3022_MOESM1_ESM.docx (477 kb)
ESM 1 (DOCX 477 kb)


  1. 1.
    Liu X, Xiang JJ, Tang Y, Zhang XL, Fu QQ, Zou JH, Lin YH (2012) Colloidal gold nanoparticle probe-based immunochromatographic assay for the rapid detection of chromium ions in water and serum samples. Anal Chim Acta 745:99–105CrossRefGoogle Scholar
  2. 2.
    Mei ZL, Deng Y, Chu HQ, Xue F, Zhong YH, Wu JJ, Yang H, Wang ZC, Zheng L, Chen W (2013) Immunochromatographic lateral flow strip for on-site detection of bisphenol A. Microchim Acta 180:279–285CrossRefGoogle Scholar
  3. 3.
    Singh J, Sharma S, Nara S (2015) Evaluation of gold nanoparticle based lateral flow assays for diagnosis of enterobacteriaceae members in food and water. Food Chem 170:470–483CrossRefGoogle Scholar
  4. 4.
    Yang QH, Gong XQ, Song T, Yang JM, Zhu SJ, Li YH, Cui Y, Li YX, Zhang BB, Chang J (2011) Quantum dot-based immunochromatography test strip for rapid, quantitative and sensitive detection of alpha fetoprotein. Biosens Bioelectron 30:145–150CrossRefGoogle Scholar
  5. 5.
    Huang XL, Aguilar ZP, Li HM, Lai WH, Wei H, Xu HY, Xiong YH (2013) Fluorescent Ru(phen)3 (2+)-doped silica nanoparticles-based ICTS sensor for quantitative detection of enrofloxacin residues in chicken meatA. Anal Chem 85:5120–5128CrossRefGoogle Scholar
  6. 6.
    Wang LY, Yan RX, Hao ZY, Wang L, Zeng JH, Bao J, Wang X, Peng Q, Li YD (2010) Fluorescence resonant energy transfer biosensor based on upconversion-luminescent nanoparticles. Angew Chem Int Ed 117:6208–6211CrossRefGoogle Scholar
  7. 7.
    Zhou J, Liu Q, Feng W, Sun Y, Li F (2015) Upconversion luminescent materials: advances and applications. Chem Rev 115:395–465CrossRefGoogle Scholar
  8. 8.
    Zhou B, Shi B, Jin D, Liu XG (2015) Controlling upconversion nanocrystals for emerging applications. Nat Nanotechnol 10:924–936CrossRefGoogle Scholar
  9. 9.
    Zhao P, Wu YY, Zhu YH, Yang XL, Jiang X, Xiao JF, Zhang YX, Li CZ (2014) Upconversion fluorescent strip sensor for rapid determination of vibrio anguillarum. Nanoscale 6:3804–3809CrossRefGoogle Scholar
  10. 10.
    Ellington AD, Szostak JW (1990) Invitro selection of RNA molecules that bind specific ligands. Nature 346:818–822CrossRefGoogle Scholar
  11. 11.
    Tuerk C, Gold L (1990) Systematic evolution of ligands by exponential enrichment-RNA ligands to bacteriophage-T4 DNA-polymerase. Science 249:505–510CrossRefGoogle Scholar
  12. 12.
    Nimjee SM, Rusconi CP, Sullenger BA (2005) Aptamers: an emerging class of therapeutics. Annu Rev Med 56:555–583CrossRefGoogle Scholar
  13. 13.
    Xu H, Mao X, Zeng QX, Wang SF, Kawde AN, Liu GD (2009) Aptamer-functionalized gold nanoparticles as probes in a dry-reagent strip biosensor for protein analysis. Anal Chem 81:669–675CrossRefGoogle Scholar
  14. 14.
    Liu GD, Mao X, Phillips JA, Xu H, Tan WH, Zeng LW (2009) Aptamer-nanoparticle strip biosensor for sensitive detection of cancer cells. Anal Chem 81:10013–10018CrossRefGoogle Scholar
  15. 15.
    Rhouat A, Yang C, Hayat A, Marty JL (2013) Aptamers: a promosing tool for ochratoxin a detection in food analysis. Toxins 5:1988–2008CrossRefGoogle Scholar
  16. 16.
    Cruzaguado JA, Penner G (2008) Determination of ochratoxin a with a DNA aptamer. J Agric Food Chem 56:10456–10461CrossRefGoogle Scholar
  17. 17.
    Zhou W, Kong W, Dou X, Zhao M, Ouyang Z, Yang M (2016) An aptamer based lateral flow strip for on-site rapid detection of ochratoxin a in astragalus membranaceus. J Chromatogr B 1022:102–108CrossRefGoogle Scholar
  18. 18.
    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 23:291–303CrossRefGoogle Scholar
  19. 19.
    Dai SL, Wu SJ, Duan N, Chen J, Zheng ZG, Wang ZP (2017) An ultrasensitive aptasensor for Ochratoxin A using hexagonal core/shell upconversion nanoparticles as luminophores. Biosens Bioelectron 91:538–544CrossRefGoogle Scholar
  20. 20.
    Wu SJ, Duan N, Zhu CQ, Ma XY, Wang M, Wang ZP (2011) Magnetic nanobead-based immunoassay for the simultaneous detection of aflatoxin B1 and ochratoxin A using upconversion nanoparticles as multicolor labels. Biosens Bioelectron 30:35–42CrossRefGoogle Scholar
  21. 21.
    Wu SJ, Duan N, Wang ZP, Wang HX (2011) Aptamer-functionalized magnetic nanoparticle-based bioassay for the detection of ochratoxin a using upconversion nanoparticles as labels. Analyst 136:2306–2314CrossRefGoogle Scholar
  22. 22.
    Dai SL, Wu SJ, Duan N, Wang ZP (2016) A luminescence resonance energy transfer based aptasensor for the mycotoxin Ochratoxin a using upconversion nanoparticles and gold nanorods. Microchim Acta 183:1909–1916CrossRefGoogle Scholar
  23. 23.
    Wu SJ, Duan N, Ma XY, Xia Y, Wang HX, Wang ZP (2013) A highly sensitive fluorescence resonance energy transfer aptasensor for staphylococcal enterotoxin B detection based on exonuclease-catalysed target recycling strategy. Anal Chim Acta 782:59–66CrossRefGoogle Scholar
  24. 24.
    Wang LB, Ma WW, Chen W, Liu LQ, Ma W, Zhu YY, Xu LG, Kuang H, Xu CL (2011) An aptamer-based chromatographic strip assay for sensitive toxin semi-quantitative detection. Biosens Bioelectron 26:3059–3062CrossRefGoogle Scholar
  25. 25.
    Zhao X, Yuan Y, Zhang X, Yue T (2014) Identification of ochratoxin a in Chinese spices using HPLC fluorescent detectors with immunoaffinity column cleanup. Food Control 46:332–337CrossRefGoogle Scholar
  26. 26.
    Wei RW, Qiu F, Kong WJ, Wei JH, Yang MH, Luo ZL, Qin JP, Ma XJ (2013) Co-occurrence of aflatoxin B1, B2, G1, G2 and ochrotoxin a in glycyrrhiza uralensis analyzed by HPLC-MS/MS. Food Control 32:216–221CrossRefGoogle Scholar
  27. 27.
    Radi AE, Muñoz-Berbel X, Cortina-Puig M, Marty JL (2009) An electrochemical immunosensor for ochratoxin a based on immobilization of antibodies on diazonium-functionalized gold electrode. Electrochim Acta 54:2180–2184CrossRefGoogle Scholar
  28. 28.
    Wu H, Liu RJ, Kang XJ, Liang CY, Lv L, Guo ZJ (2018) Fluorometric aptamer assay for ochratoxin a based on the use of single walled carbon nanohorns and exonuclease III-aided amplification. Microchim Acta 185:27CrossRefGoogle Scholar
  29. 29.
    Chen JH, Fang ZY, Liu J, Zeng LW (2012) A simple and rapid biosensor for ochratoxin a based on a structure-switching signaling aptamer. Food Control 25:555–560CrossRefGoogle Scholar
  30. 30.
    Yu XH, Lin YH, Wang XS, Xu LJ, Wang ZW, Fu FF (2018) Exonuclease-assisted multicolor aptasensor for visual detection of ochratoxin a based on G-quadruplex-hemin DNAzyme-mediated etching of gold nanorods. Microchim Acta 185:259CrossRefGoogle Scholar
  31. 31.
    Wang CQ, Qian J, Wang K, Yang XW, Liu Q, Hao N, Wang CK, Dong XY, Huang XY (2016) Colorimetric aptasensing of ochratoxin a using au@Fe3O4 nanoparticles as signal indicator and magnetic separator. Biosens Bioelectron 77:1183–1191CrossRefGoogle Scholar
  32. 32.
    Laura A, Gilda D, Claudio B, Cristina G, Gianfranco G (2011) A lateral flow immunoassay for measuring ochratoxin a: development of a single system for maize, wheat and durum wheat. Food Control 22:1965–1970CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.State Key Laboratory of Food Science and TechnologyJiangnan UniversityWuxiChina
  2. 2.School of Food Science and TechnologyJiangnan UniversityWuxiChina
  3. 3.School of Food and Biological EngineeringJiangsu UniversityZhenjiangChina
  4. 4.China Rural Technology Development CenterBeijingChina
  5. 5.School of Food and Chemical EngineeringBeijing Technology and Business UniversityBeijingChina

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