Skip to main content
SpringerLink
Log in

Simultaneous determination for A. flavus–metabolizing mycotoxins by time-resolved fluorescent microbead or gold-enabling test strip in agricultural products based on monoclonal antibodies

  • Original Paper
  • Published:
Microchimica Acta Aims and scope Submit manuscript

Abstract

To simultaneously detect two metabolites of Aspergillus flavus, namely, cyclopiazonic acid (CPA) and aflatoxin (AFT), an ultrasensitive monoclonal antibody (mAb) YTT-2 against CPA was developed and characterized, with sensitivity to CPA of 1.32 ng mL−1. Along with the previously homemade mAb 1C11 against AFT, two mAbs were used to develop time-resolved fluorescence immunoprobes or gold immunoprobes. We developed two multiple-analyte paper immunosensors including time-resolved fluorescent immunochromatographic assay (TRFICA) and gold immunochromatographic assay (GICA) for the simultaneous determination of CPA and AFT. The TRFICA showed limits of determination (LODs) of 0.21 and 0.004 ng mL−1, while the GICA showed LODs of 0.33 and 0.01 ng mL−1 for CPA and AFT, respectively. To validate the specificity of the two rapid immunoassays, rice, corn and peanut samples were spiked with different concentrations of CPA and AFT. The two methods showed satisfactory recoveries (76.39~90.82% for CPA and 84.60~94.45% for AFT) and coefficients of variation of 3.50~7.80% for CPA and 4.12~13.90% for AFT. The results indicated that the TRFICA could complete the test within 5 min and had lower LODs and linear ranges, compared with that of GICA. The method developed in this work can be widely applied to the rapid and quantitative simultaneous determination of multiple harmful metabolites in fungi for food safety and health care.

Graphical abstract

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Heperkan D, Karbancioglu FG, Oktay HI (2012) Mycoflora and natural occurrence of aflatoxin, cyclopiazonic acid, fumonisin and ochratoxin A in dried figs. Food Addit Contam 29(2):277–286

    Article  CAS  Google Scholar 

  2. Yazdanpanah H (2011) Mycotoxins: analytical challenges. Iranian J Pharmaceut Res Ijpr 10(4):653

    Google Scholar 

  3. Oliveira CAF, Sebastião LS, Fagundes H, Rosim RE, Fernandes AM (2008) Aflatoxins and cyclopiazonic acid in feed and milk from dairy farms in São Paulo, Brazil. Food Addit Contam Part B Surveill 1(2):147–152

    Article  CAS  Google Scholar 

  4. Georgianna DR, Fedorova ND, Burroughs JL, Dolezal AL, Bok JW, Horowitz-Brown S, Woloshuk CP, Yu JJ, Keller NP, Payne GA (2010) Beyond aflatoxin: four distinct expression patterns and functional roles associated with Aspergillus flavus secondary metabolism gene clusters. Mol Plant Pathol 11(2):213–226

    Article  CAS  Google Scholar 

  5. Tokuoka M, Seshime Y, Fujii I, Kitamoto K, Takahashi T, Koyama Y (2008) Identification of a novel polyketide synthase–nonribosomal peptide synthetase (PKS–NRPS) gene required for the biosynthesis of cyclopiazonic acid in Aspergillus oryzae. Fungal Genet Biol 45:1608–1615

    Article  CAS  Google Scholar 

  6. Zorzete P, Baquiao AC, Atayde DD, Reis TA, Goncalez E, Correa B (2013) Mycobiota, aflatoxins and cyclopiazonic acid in stored peanut cultivars. Food Res Int 52(1):380–386

    Article  CAS  Google Scholar 

  7. Heperkan D, Somuncuoglu S, Karbancioglu-Güler F, Mecik N (2012) Natural contamination of cyclopiazonic acid in dried figs and co-occurrence of aflatoxin. Food Control 23(1):82–86

    Article  CAS  Google Scholar 

  8. Lansden JA (1986) Determination of cyclopiazonic acid in peanuts and corn by thin layer chromatography. JAOAC 69(6):964–966

    CAS  Google Scholar 

  9. Hayashi Y, Yoshizawa T (2005) Analysis of cyclopiazonic acid in corn and rice by a newly developed method. Food Chem 93(2):215–221

    Article  CAS  Google Scholar 

  10. Ansari P, Häubl G (2016) Determination of cyclopiazonic acid in white mould cheese by liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) using a novel internal standard. Food Chem 211:978–982

    Article  CAS  Google Scholar 

  11. Tang X, Li P, Zhang Z, Zhang Q, Guo J, Zhang W (2017) An ultrasensitive gray-imaging-based quantitative immunochromatographic detection method for fumonisin B1 in agricultural products. Food Control 80:333–340

  12. Vdovenko MM, Lu CC, Yu FY, Sakharov IY (2014) Development of ultrasensitive direct chemiluminescent enzyme immunoassay for determination of aflatoxin M1 in milk. Food Chem 158:310–314

    Article  CAS  Google Scholar 

  13. Wang D, Zhang Z, Li P, Zhang Q, Ding X, Zhang W (2015) Europium nanospheres-based time-resolved fluorescence for rapid and ultrasensitive determination of total aflatoxin in feed. J Agric Food Chem 63:10313–10318

    Article  CAS  Google Scholar 

  14. Zhang D, Li P, Zhang Q, Zhang W (2011) Ultrasensitive nanogold probe-based immunochromatographic assay for simultaneous detection of total aflatoxins in peanuts. Biosens Bioelectron 26(6):2877–2882

    Article  CAS  Google Scholar 

  15. Zhang D, Li P, Zhang Q, Zhang W, Huang Y, Ding X, Jiang J (2009) Production of ultrasensitive generic monoclonal antibodies against major aflatoxins using a modified two-step screening procedure. Anal Chim Acta 636(1):63–69

    Article  CAS  Google Scholar 

  16. Hahnau S, Weiler EW (1993) Monoclonal antibodies for the enzyme immunoassay of the mycotoxin cyclopiazonic acid. J Agric Food Chem 41(7):1076–1080

    Article  CAS  Google Scholar 

  17. Li P, Qi Z, Wen Z, Zhang J, Chen X, Jiang J, Xie L, Zhang D (2009) Development of a class-specific monoclonal antibody-based ELISA for aflatoxins in peanut. Food Chem 115(1):313–317

    Article  CAS  Google Scholar 

  18. Majdinasab M, Sheikh-Zeinoddin M, Soleimanian-Zad S, Li PW, Zhang Q, Li X, Tang XQ, Li J (2015) A reliable and sensitive time-resolved fluorescent immunochromatographic assay (TRFICA) for ochratoxin A in agro-products. Food Control 47:126–134

    Article  CAS  Google Scholar 

  19. Yang Q, Zhu J, Ma F, Li P, Zhang L, Zhang W, Ding X, Zhang Q (2016) Quantitative determination of major capsaicinoids in serum by ELISA and time-resolved fluorescent immunoassay based on monoclonal antibodies. Biosens Bioelectron 81:229–235

    Article  CAS  Google Scholar 

  20. Astoreca A, Vaamonde G, Dalcero A, Marin S, Ramos A (2014) Abiotic factors and their interactions influence on the co-production of aflatoxin B(1) and cyclopiazonic acid by Aspergillus flavus isolated from corn. Food Microbiol 38:276–283

    Article  CAS  Google Scholar 

  21. Hahnau S, Weiler EW (1991) Determination of the mycotoxin cyclopiazonic acid by enzyme immunoassay. J Agric Food Chem 39(10):1887–1891

    Article  CAS  Google Scholar 

  22. Li Y, Liu L, Kuang H, Xu C (2020) Visible and eco-friendly immunoassays for the detection of cyclopiazonic acid in maize and rice. J Food Sci 85(1):105–113

    Article  CAS  Google Scholar 

  23. Maragos CM, Sieve KK, Bobell J (2017) Detection of cyclopiazonic acid (CPA) in maize by immunoassay. Mycotoxin Res 33(2):157–165

    Article  CAS  Google Scholar 

Download references

Funding

This work was supported by the National Key Research and Development Program of China (2018YFC1602500), the Natural Science Foundation of China (32072331), and Agricultural Science and Technology Innovation Program of CAAS (CAAS- ZDRW202011).

Author information

Authors and Affiliations

  1. Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan, 430062, China

    Tingting Yan, Zhaowei Zhang, Qi Zhang, Xiaoqian Tang, Du Wang, Xiaofeng Hu, Wen Zhang, Xiaomei Chen & Peiwu Li

  2. Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture and Rural Affairs, Wuhan, 430062, China

    Tingting Yan, Zhaowei Zhang, Qi Zhang, Xiaoqian Tang, Du Wang, Xiaofeng Hu, Wen Zhang, Xiaomei Chen & Peiwu Li

  3. Quality Inspection and Test Center for Oilseeds Products, Ministry of Agriculture and Rural Affairs, Wuhan, 430062, China

    Qi Zhang, Xiaomei Chen & Peiwu Li

  4. Laboratory of Quality and Safety Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture and Rural Affairs, Wuhan, 430062, China

    Wen Zhang & Peiwu Li

  5. Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture and Rural Affairs, Wuhan, 430062, China

    Peiwu Li

Authors
  1. Tingting Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhaowei Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Qi Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xiaoqian Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Du Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xiaofeng Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Wen Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xiaomei Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Peiwu Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Zhaowei Zhang, Qi Zhang or Peiwu Li.

Ethics declarations

Conflict of interest

The authors declare that they have no conflicts of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

ESM 1

(DOCX 400 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yan, T., Zhang, Z., Zhang, Q. et al. Simultaneous determination for A. flavus–metabolizing mycotoxins by time-resolved fluorescent microbead or gold-enabling test strip in agricultural products based on monoclonal antibodies. Microchim Acta 187, 653 (2020). https://doi.org/10.1007/s00604-020-04623-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s00604-020-04623-x

Keywords

Search

Navigation