Advertisement

GO-amplified fluorescence polarization assay for high-sensitivity detection of aflatoxin B1 with low dosage aptamer probe

  • Hua Ye
  • Qianqian Lu
  • Nuo Duan
  • Zhouping WangEmail author
Research Paper

Abstract

Aflatoxin B1 (AFB1) is the most toxic mycotoxin of the aflatoxins (AFs) and shows carcinogenic, teratogenic and mutagenic effects in humans and animals. AFB1 is widely seen in cereal products such as rice and wheat. This research proposed a low-cost, high-sensitivity fluorescence polarization (FP) assay for detection of AFB1 using aptamer biosensors based on graphene oxide (GO). The aptamers labelled with fluorescein amidite (FAM) were adsorbed on the surface of GO through π–π stacking and electrostatic interaction, thus forming aptamer/GO macromolecular complexes. Under these conditions, the local rotation of fluorophores was limited and the system had a high FP value. When there was AFB1 in the system, aptamers were dissociated from the GO surface and combined with AFB1 owing to their specificity to form aptamer/AFB1 complexes. As a result, large changes were observed in the molecular weights of aptamers before, and after, the combination, therefore leading to the apparent changes in FP value. The results showed that when only 10 nM of aptamer was used, the changes in FP and the AFB1 concentration had a favourable linear relationship within 0.05 to 5 nM of AFB1, and the lowest detection limit (LOD) was 0.05 nM. In addition, the recoveries of rice sample extract ranged from 89.2% to 112%. The method is simple, highly sensitive, cost-efficient and shows potential application prospects.

Keywords

Aptamer Fluorescence polarization Graphene oxide Signal amplifier Aflatoxin B1 

Notes

Acknowledgements

This work was partly funded by the Science & Technology Support Program of Jiangsu Province [BE2017623], Jiangsu Agricultural Science and Technology Innovation Fund [CX(18)2025], the National Natural Science Fund of China [31871881], the Fundamental Research Funds for Central Universities [JUSRP51714B], the Distinguished Professor Program of Jiangsu Province and Jiangnan University Postgraduate Overseas Research Project.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human or animal subjects.

Supplementary material

216_2018_1540_MOESM1_ESM.pdf (218 kb)
ESM 1 (PDF 218 kb)

References

  1. 1.
    Shephard GS. Aflatoxin analysis at the beginning of the twenty-first century. Anal Bioanal Chem. 2009;395(5):1215–24.  https://doi.org/10.1007/s00216-009-2857-y.CrossRefPubMedGoogle Scholar
  2. 2.
    Marin S, Ramos AJ, Cano-Sancho G, Sanchis V. Mycotoxins: occurrence, toxicology, and exposure assessment. Food Chem Toxicol. 2013;60:218–37.  https://doi.org/10.1016/j.fct.2013.07.047. CrossRefPubMedGoogle Scholar
  3. 3.
    Verheecke C, Liboz T, Mathieu F. Microbial degradation of aflatoxin B1: current status and future advances. Int J Food Microbiol. 2016;237:1–9.  https://doi.org/10.1016/j.ijfoodmicro.2016.07.028.CrossRefPubMedGoogle Scholar
  4. 4.
    Henry SH, Bosch FX, Troxell TC, Bolger PM. Reducing liver cancer–global control of aflatoxin. Science. 1999;286(5449):2453–4.  https://doi.org/10.1126/science.286.5449.2453.
  5. 5.
    Geleta GS, Zhao Z, Wang Z. A novel reduced graphene oxide/molybdenum disulfide/polyaniline nanocomposite-based electrochemical aptasensor for detection of aflatoxin B1. Analyst. 2018;143(7):1644–9.  https://doi.org/10.1039/c7an02050c.CrossRefPubMedGoogle Scholar
  6. 6.
    Sartori AV, de Moraes MHP, dos Santos RP, Souza YP, da Nóbrega AW. Determination of mycotoxins in cereal-based porridge destined for infant consumption by ultra-high performance liquid chromatography-tandem mass spectrometry. Food Anal Methods. 2017;10(12):4049–61.  https://doi.org/10.1007/s12161-017-0965-4.CrossRefGoogle Scholar
  7. 7.
    Aiko V, Mehta A. Occurrence, detection and detoxification of mycotoxins. J Biosci. 2015;40(5):943–54.  https://doi.org/10.1007/s12038-015-9569-6.CrossRefPubMedGoogle Scholar
  8. 8.
    Donna L, Orti JG, Ashley DL, Hill RH Jr. Chromatographic and spectroscopic properties of hemiacetals of aflatoxin and sterigmatocystin metabolites. J Chromatogr A. 1989;462:269–79.CrossRefGoogle Scholar
  9. 9.
    Zdena Ďuračková VB, Nemec P. Systematic analysis of mycotoxins by thin-layer chromatography. J Chromatogr A. 1976;116(1):141–54.CrossRefGoogle Scholar
  10. 10.
    Alizadeh N, Memar MY, Mehramuz B, Abibiglou SS, Hemmati F, Samadi Kafil H. Current advances in aptamer-assisted technologies for detecting bacterial and fungal toxins. J Appl Microbiol. 2018;124(3):644–51.  https://doi.org/10.1111/jam.13650.CrossRefPubMedGoogle Scholar
  11. 11.
    Kolosova AY, Shim WB, Yang ZY, Eremin SA, Chung DH. Direct competitive ELISA based on a monoclonal antibody for detection of aflatoxin B1. Stabilization of ELISA kit components and application to grain samples. Anal Bioanal Chem. 2006;384(1):286–94.  https://doi.org/10.1007/s00216-005-0103-9.CrossRefPubMedGoogle Scholar
  12. 12.
    Anfossi L, Baggiani C, Giovannoli C, D’Arco G, Giraudi G. Lateral-flow immunoassays for mycotoxins and phycotoxins: a review. Anal Bioanal Chem. 2013;405(2–3):467–80.  https://doi.org/10.1007/s00216-012-6033-4.CrossRefPubMedGoogle Scholar
  13. 13.
    Jiang W, Wang Z, Nölke G, Zhang J, Niu L, Shen J. Simultaneous determination of aflatoxin B1 and aflatoxin M1 in food matrices by enzyme-linked immunosorbent assay. Food Anal Methods. 2012;6(3):767–74.  https://doi.org/10.1007/s12161-012-9484-5.CrossRefGoogle Scholar
  14. 14.
    Li J, Fang X, Yang Y, Cheng X, Tang P. An improved chemiluminescence immunoassay for the ultrasensitive detection of aflatoxin B1. Food Anal Methods. 2016;9(11):3080–6.  https://doi.org/10.1007/s12161-016-0499-1.CrossRefGoogle Scholar
  15. 15.
    Yu X, Li Z, Zhao M, Lau SCS, Ru Tan H, Teh WJ, et al. Quantification of aflatoxin B1 in vegetable oils using low temperature clean-up followed by immuno-magnetic solid phase extraction. Food Chem. 2018.  https://doi.org/10.1016/j.foodchem.2018.09.132.
  16. 16.
    Mairal T, Ozalp VC, Lozano Sanchez P, Mir M, Katakis I, O’Sullivan CK. Aptamers: molecular tools for analytical applications. Anal Bioanal Chem. 2008;390(4):989–1007.  https://doi.org/10.1007/s00216-007-1346-4.CrossRefPubMedGoogle Scholar
  17. 17.
    Le Chryseis L, Cruz-Aguado JA, Penner GA. DNA ligands for aflatoxin and zearalenone. USA patent PTC/CA2010/001292. 2012.Google Scholar
  18. 18.
    Shim W-B, Mun H, Joung H-A, Ofori JA, Chung D-H, Kim M-G. Chemiluminescence competitive aptamer assay for the detection of aflatoxin B1 in corn samples. Food Control. 2014;36(1):30–5.  https://doi.org/10.1016/j.foodcont.2013.07.042.CrossRefGoogle Scholar
  19. 19.
    Ma X, Wang W, Chen X, Xia Y, Wu S, Duan N, et al. Selection, identification, and application of aflatoxin B1 aptamer. Eur Food Res Technol. 2014;238(6):919–25.  https://doi.org/10.1007/s00217-014-2176-1.CrossRefGoogle Scholar
  20. 20.
    Setlem K, Mondal B, Ramlal S, Kingston J. Immuno affinity SELEX for simple, rapid, and cost-effective aptamer enrichment and identification against aflatoxin B1. Front Microbiol. 2016;7:1909.  https://doi.org/10.3389/fmicb.2016.01909.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Aswani Kumar YVV, Renuka RM, Achuth J, Venkataramana M, Ushakiranmayi M, Sudhakar P. Development of hybrid IgG-aptamer sandwich immunoassay platform for aflatoxin B1 detection and its evaluation onto various field samples. Front Pharmacol. 2018;9:271.  https://doi.org/10.3389/fphar.2018.00271.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Jafari M, Rezaei M, Kalantari H, Tabarzad M, Daraei B. DNAzyme-aptamer or aptamer-DNAzyme paradigm: biochemical approach for aflatoxin analysis. Biotechnol Appl Biochem. 2018;65(2):274–80.  https://doi.org/10.1002/bab.1563.CrossRefPubMedGoogle Scholar
  23. 23.
    Li Y, Sun L, Zhao Q. Development of aptamer fluorescent switch assay for aflatoxin B1 by using fluorescein-labeled aptamer and black hole quencher 1-labeled complementary DNA. Anal Bioanal Chem. 2018.  https://doi.org/10.1007/s00216-018-1237-x.
  24. 24.
    Peng G, Li X, Cui F, Qiu Q, Chen X, Huang H. Aflatoxin B1 electrochemical aptasensor based on tetrahedral DNA nanostructures functionalized three dimensionally ordered macroporous MoS2-AuNPs film. ACS Appl Mater Interfaces. 2018;10(21):17551–9.  https://doi.org/10.1021/acsami.8b01693.CrossRefPubMedGoogle Scholar
  25. 25.
    Qian J, Ren C, Wang C, Chen W, Lu X, Li H, et al. Magnetically controlled fluorescence aptasensor for simultaneous determination of ochratoxin A and aflatoxin B1. Anal Chim Acta. 2018;1019:119–27.  https://doi.org/10.1016/j.aca.2018.02.063.CrossRefPubMedGoogle Scholar
  26. 26.
    Sun L, Zhao Q. Competitive horseradish peroxidase-linked aptamer assay for sensitive detection of aflatoxin B1. Talanta. 2018;179:344–9.  https://doi.org/10.1016/j.talanta.2017.11.048.CrossRefPubMedGoogle Scholar
  27. 27.
    Wu W, Zhu Z, Li B, Liu Z, Jia L, Zuo L, et al. A direct determination of AFBs in vinegar by aptamer-based surface plasmon resonance biosensor. Toxicon. 2018;146:24–30.  https://doi.org/10.1016/j.toxicon.2018.03.006.CrossRefPubMedGoogle Scholar
  28. 28.
    Zhang C, Dou X, Zhang L, Sun M, Zhao M, OuYang Z, et al. A rapid label-free fluorescent aptasensor PicoGreen-based strategy for aflatoxin B(1) detection in traditional Chinese medicines. Toxins (Basel). 2018;10(3).  https://doi.org/10.3390/toxins10030101.
  29. 29.
    Zhu C, Zhang G, Huang Y, Yang S, Ren S, Gao Z, et al. Dual-competitive lateral flow aptasensor for detection of aflatoxin B1 in food and feedstuffs. J Hazard Mater. 2018;344:249–57.  https://doi.org/10.1016/j.jhazmat.2017.10.026.CrossRefPubMedGoogle Scholar
  30. 30.
    Zhang J, Li Z, Zhao S, Lu Y. Size-dependent modulation of graphene oxide-aptamer interactions for an amplified fluorescence-based detection of aflatoxin B1 with a tunable dynamic range. Analyst. 2016;141(13):4029–34.  https://doi.org/10.1039/c6an00368k.CrossRefPubMedGoogle Scholar
  31. 31.
    Joo M, Baek SH, Cheon SA, Chun HS, Choi S-W, Park TJ. Development of aflatoxin B1 aptasensor based on wide-range fluorescence detection using graphene oxide quencher. Colloids Surf B Biointerfaces. 2017;154:27–32.  https://doi.org/10.1016/j.colsurfb.2017.03.010.CrossRefPubMedGoogle Scholar
  32. 32.
    Smith DS, Eremin SA. Fluorescence polarization immunoassays and related methods for simple, high-throughput screening of small molecules. Anal Bioanal Chem. 2008;391(5):1499–507.  https://doi.org/10.1007/s00216-008-1897-z.CrossRefPubMedGoogle Scholar
  33. 33.
    Mohammad Sarwar Nasir MEJ. Development of a fluorescence polarization assay for the determination of aflatoxins in grains. J Agric Food Chem. 2002;50(11):3116–21.CrossRefGoogle Scholar
  34. 34.
    Sheng YJ, Eremin S, Mi TJ, Zhang SX, Shen JZ, Wang ZH. The development of a fluorescence polarization immunoassay for aflatoxin detection. Biomed Environ Sci. 2014;27(2):126–9.  https://doi.org/10.3967/bes2014.027. CrossRefPubMedGoogle Scholar
  35. 35.
    Beloglazova NV, Eremin SA. Rapid screening of aflatoxin B1 in beer by fluorescence polarization immunoassay. Talanta. 2015;142:170–5.  https://doi.org/10.1016/j.talanta.2015.04.027.CrossRefPubMedGoogle Scholar
  36. 36.
    Huang H, Qin J, Hu K, Liu X, Zhao S, Huang Y. Novel autonomous protein-encoded aptamer nanomachines and isothermal exponential amplification for ultrasensitive fluorescence polarization sensing of small molecules. RSC Adv. 2016;6(89):86043–50.  https://doi.org/10.1039/c6ra17959b.CrossRefGoogle Scholar
  37. 37.
    Sun L, Zhao Q. Direct fluorescence anisotropy approach for aflatoxin B1 detection and affinity binding study by using single tetramethylrhodamine labeled aptamer. Talanta. 2018;189:442–50.  https://doi.org/10.1016/j.talanta.2018.07.036.CrossRefPubMedGoogle Scholar
  38. 38.
    Yu Y, Liu Y, Zhen SJ, Huang CZ. A graphene oxide enhanced fluorescence anisotropy strategy for DNAzyme-based assay of metal ions. Chem Commun (Camb). 2013;49(19):1942–4.  https://doi.org/10.1039/c3cc38129c.CrossRefGoogle Scholar
  39. 39.
    Li X, Ding X, Li Y, Wang L, Fan J. A TiS2 nanosheet enhanced fluorescence polarization biosensor for ultra-sensitive detection of biomolecules. Nanoscale. 2016;8(18):9852–60.  https://doi.org/10.1039/c6nr00946h.CrossRefPubMedGoogle Scholar
  40. 40.
    Chen Z, Li H, Jia W, Liu X, Li Z, Wen F, et al. Bivalent aptasensor based on silver-enhanced fluorescence polarization for rapid detection of lactoferrin in milk. Anal Chem. 2017;89(11):5900–8.  https://doi.org/10.1021/acs.analchem.7b00261. CrossRefPubMedGoogle Scholar
  41. 41.
    Liu J, Wang C, Jiang Y, Hu Y, Li J, Yang S, et al. Graphene signal amplification for sensitive and real-time fluorescence anisotropy detection of small molecules. Anal Chem. 2013;85(3):1424–30.  https://doi.org/10.1021/ac3023982. CrossRefPubMedGoogle Scholar
  42. 42.
    Xiao X, Li YF, Huang CZ, Zhen SJ. A novel graphene oxide amplified fluorescence anisotropy assay with improved accuracy and sensitivity. Chem Commun (Camb). 2015;51(89):16080–3.  https://doi.org/10.1039/c5cc05902j.CrossRefGoogle Scholar
  43. 43.
    Zhen SJ, Xiao X, Li CH, Huang CZ. An enzyme-free DNA circuit-assisted graphene oxide enhanced fluorescence anisotropy assay for microRNA detection with improved sensitivity and selectivity. Anal Chem. 2017;89(17):8766–71.  https://doi.org/10.1021/acs.analchem.7b00955.CrossRefPubMedGoogle Scholar
  44. 44.
    Cui L, Zou Y, Lin N, Zhu Z, Jenkins G, Yang CJ. Mass amplifying probe for sensitive fluorescence anisotropy detection of small molecules in complex biological samples. Anal Chem. 2012;84(13):5535–41.  https://doi.org/10.1021/ac300182w.CrossRefPubMedGoogle Scholar
  45. 45.
    Du Y, Moulick K, Rodina A, Aguirre J, Felts S, Dingledine R, et al. High-throughput screening fluorescence polarization assay for tumor-specific Hsp90. J Biomol Screen. 2007;12(7):915–24.  https://doi.org/10.1177/1087057107306067.CrossRefPubMedGoogle Scholar
  46. 46.
    Huang Y, Liu X, Zhang L, Hu K, Zhao S, Fang B, et al. Nicking enzyme and graphene oxide-based dual signal amplification for ultrasensitive aptamer-based fluorescence polarization assays. Biosens Bioelectron. 2015;63:178–84.  https://doi.org/10.1016/j.bios.2014.07.036.CrossRefPubMedGoogle Scholar
  47. 47.
    Aswani Kumar YVV, Renuka RM, Achuth J, Mudili V, Poda S. Development of a FRET-based fluorescence aptasensor for the detection of aflatoxin B1 in contaminated food grain samples. RSC Adv. 2018;8(19):10465–73.  https://doi.org/10.1039/c8ra00317c. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Hua Ye
    • 1
    • 2
  • Qianqian Lu
    • 1
  • Nuo Duan
    • 1
  • Zhouping Wang
    • 1
    • 3
    • 4
    • 5
    Email author
  1. 1.State Key Laboratory of Food Science and Technology, School of Food Science and TechnologyJiangnan UniversityWuxiChina
  2. 2.College of Food EngineeringAnhui Science and Technology UniversityFengyangChina
  3. 3.National Engineering Research Center of Seafood, School of Food Science and TechnologyDalian Polytechnic UniversityDalianChina
  4. 4.International Joint Laboratory on Food SafetyJiangnan UniversityWuxiChina
  5. 5.Collaborative Innovation Center of Food Safety and Quality Control of Jiangsu ProvinceWuxiChina

Personalised recommendations