Microchimica Acta

, Volume 180, Issue 13–14, pp 1217–1223 | Cite as

Highly sensitive FRET-based fluorescence immunoassay for aflatoxin B1 using cadmium telluride quantum dots

  • Roya Zekavati
  • Shahabeddin Safi
  • Seyed Jamal Hashemi
  • Tavoos Rahmani-Cherati
  • Meisam Tabatabaei
  • Afshin MohsenifarEmail author
  • Mansour BayatEmail author
Original Paper


We report on a competitive immunoassay for the determination of aflatoxin B1 using fluorescence resonance energy transfer (FRET) from anti-aflatoxin B1 antibody (immobilized on the shell of CdTe quantum dots) to Rhodamine 123 (Rho 123-labeled aflatoxin B1 bound to albumin). The highly specific immunoreaction between the antibody against aflatoxin B1 on the QDs and the labeled-aflatoxin B1 brings the Rho 123 fluorophore (acting as the acceptor) and the QDs (acting as the donor) in close spatial proximity and causes FRET to occur upon photoexcitation of the QDs. In the absence of unlabeled aflatoxin B1, the antigen-antibody complex is stable, and strong emission resulting from the FRET from QDs to labeled aflatoxin B1 is observed. In the presence of aflatoxin B1, it will compete with the labeled aflatoxin B1-albumin complex for binding to the antibody-QDs conjugate so that FRET will be increasingly suppressed. The reduction in the fluorescence intensity of the acceptor correlates well with the concentration of aflatoxin B1. The feasibility of the method was established by the detection of aflatoxin B1 in spiked human serum. There is a linear relationship between the increased fluorescence intensity of Rho 123 with increasing concentration of aflatoxin B1 in spike human serum, over the range of 0.1–0.6 μmol·mL−1. The limit of detection is 2 × 10−11 M. This homogeneous competitive detection scheme is simple, rapid and efficient, and does not require excessive washing and separation steps.


A nanobiosensor has been fabricated based on a competitive immunoassay for the determination of aflatoxin B1 using fluorescence resonance energy transfer (FRET). In the presence of aflatoxin B1, it will compete with the labeled aflatoxin B1-albumin complex for binding to the antibody-QDs conjugate so that FRET will be increasingly suppressed.


Aflatoxin B1 Nanobiosensor FRET Immunoreaction Mycotoxin 



We appreciate the financial support provided by Nanozino Company. The authors would like to thank Mrs. Batool Etemadikia for her valuable technical assistance.


  1. 1.
    Devi KT, Mayo MA, Reddy KL, Delfosse P, Reddy G, Reddy SV, Reddy DV (1999) Production and characterization of monoclonal antibodies for aflatoxin B1. Lett Appl Microbiol 29:284CrossRefGoogle Scholar
  2. 2.
    Hussein HS, Brasel JM (2001) Toxicity, metabolism, and impact of mycotoxins on humans and animals. Toxicol 167:101CrossRefGoogle Scholar
  3. 3.
    Kamkar A, Jahed Khaniki GR, Alavi AS (2011) Occurrence of aflatoxin M1 in raw milk produced in Ardabil of Iran. Iran J Environ Health Sci Eng 8:123–128Google Scholar
  4. 4.
    Oldano F, Ruffier M, Sezian A, Bandirola C (2007) Aflatoxins occurrence in milk and feed in Northern Italy during 2004–2005. Food Control 18:1263CrossRefGoogle Scholar
  5. 5.
    Sapsford KE, Taitt CR, Fertig S, Moore MH, Lassman ME, Maragos CM, Shriver-Lake LC (2006) Indirect competitive immunoassay for detection of aflatoxin B1 in corn and nut products using the array biosensor. Biosens Bioelectron 21:2298CrossRefGoogle Scholar
  6. 6.
    Liao JY, Li H (2010) Lateral flow immunodipstick for visual detection of aflatoxin B1 in food using immuno-nanoparticles composed of a silver core and a gold shell. Microchim Acta 171:289CrossRefGoogle Scholar
  7. 7.
    Xiulan S, Xiaolian Z, Jian T, Xiaohong G, Jun Z, Chu FS (2006) Preparation of gold-labeled antibody probe and its use in immunochromatography assay for detection of aflatoxin B1. Int J Food Microbiol 99:185CrossRefGoogle Scholar
  8. 8.
    Xiulan S, Xiaolian Z, Jian T, Xiaohong G, Jun Z, Chu FS (2006) Development of an immunochromatographic assay for detection of aflatoxin B1 in foods. Food Control 17:256CrossRefGoogle Scholar
  9. 9.
    Lamberti I, Tanzarella C, Solinas I, Padula C, Mosiello L (2009) An antibody-based microarray assay for the simultaneous detection of aflatoxin B1 and fumonisin B1. Mycotox Res 25:193CrossRefGoogle Scholar
  10. 10.
    Bacher G, Pal S, Kanungo L, Bhand S (2012) A label-free silver wire based impedimetric immunosensor for detection of aflatoxin M1 in milk. Sensors Actuators B Chem 128:223CrossRefGoogle Scholar
  11. 11.
    Dong F, Kewang H, Han H, Liang J (2009) A novel method for methimazole determination using CdSe quantum dots as fluorescence probes. Microchim Acta 165:195CrossRefGoogle Scholar
  12. 12.
    Rad F, Mohsenifar A, Tabatabaei M, Safarnejad MR, Sahryari F, Safarpour H, Foroutan A, Mardi M, Davoudi D, Fotokian M (2012) Detection of candidatus phytoplasma aurantifolia with a quantum dots FRET-based biosensor. J Plant Pathol 94:525Google Scholar
  13. 13.
    Costa-Fernandez JM (2006) Optical sensors based on luminescent quantum dots. Anal Bioanal Chem 384:37CrossRefGoogle Scholar
  14. 14.
    Clapp AR, Medintz IL, Mattoussi H (2006) Förster resonance energy transfer investigations using quantum-dot fluorophores. Chemphyschem 16:47CrossRefGoogle Scholar
  15. 15.
    Zhong P, He G, Zhang M (2012) Optimal spectra of white light-emitting diodes using quantum dot nanophosphors. Opt Express 20:9122CrossRefGoogle Scholar
  16. 16.
    Zhang H, Liu L, Fu X, Zhu Z (2012) Microfluidic beads-based immunosensor for sensitive detection of cancer biomarker proteins using multienzyme-nanoparticle amplification and quantum dots labels. Biosens Bioelectron 42:23CrossRefGoogle Scholar
  17. 17.
    Jaiswal JK, Goldman ER, Mattoussi H, Simon SM (2004) Use of quantum dots for live cell imaging. Nat Methods 1:73CrossRefGoogle Scholar
  18. 18.
    Medintz IL, Konnert JH, Clapp AR, Stanish I, Twigg ME, Mattoussi H, Mauro JM, Deschamps JR (2004) A fluorescence resonance energy transfer-derived structure of a quantum dot-protein bioconjugate nanoassembly. Proc Natl Acad Sci U S A 101:9612CrossRefGoogle Scholar
  19. 19.
    Goldman ER, Balighian ED, Mattoussi H, Kuno MK, Mauro JM, Tran PT, Anderson GP (2002) Avidin: a natural bridge for quantum dot-antibody conjugates. J Am Chem Soc 124:6378CrossRefGoogle Scholar
  20. 20.
    Algar WR, Malanoski AP, Susumu K, Stewart MH, Hildebrandt N, Medintz IL (2012) Multiplexed tracking of protease activity using a single color of quantum dot vector and a time-gated förster resonance energy transfer relay. Anal Chem 84:10136CrossRefGoogle Scholar
  21. 21.
    Pei J, Zhu H, Wang X, Zhang H, Yang X (2012) Synthesis of cysteamine-coated CdTe quantum dots and its application in mercury (II) detection. Anal Chim Acta 757:63CrossRefGoogle Scholar
  22. 22.
    Medintz IL, Mattoussi H (2009) Quantum dot-based resonance energy transfer and its growing application in biology. Phys Chem Chem Phys 11:17CrossRefGoogle Scholar
  23. 23.
    Shamsipur M, Shanehasz M, Khajeh K, Mollania N, Kazemi SH (2012) A novel quantum dot-laccase hybrid nanobiosensor for low level determination of dopamine. Analyst 137:5553CrossRefGoogle Scholar
  24. 24.
    Shanehsaz M, Mohsenifar A, Hasannia S, Pirooznia N, Samaei Y, Shamsipur M (2013) Detection of Helicobacter pylori with a anobiosensor based on fluorescence resonance energy transfer using CdTe quantum dots. Microchim Acta 180:195CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Wien 2013

Authors and Affiliations

  • Roya Zekavati
    • 1
  • Shahabeddin Safi
    • 2
  • Seyed Jamal Hashemi
    • 3
  • Tavoos Rahmani-Cherati
    • 4
    • 6
  • Meisam Tabatabaei
    • 5
  • Afshin Mohsenifar
    • 5
    • 6
    Email author
  • Mansour Bayat
    • 1
    Email author
  1. 1.Department of Mycology, Faculty of Veterinary Specialized SciencesScience and Research BranchTehranIran
  2. 2.Department of Clinical Pathology, Faculty of Veterinary Specialized SciencesScience and Research BranchTehranIran
  3. 3.Department of Medical Mycology and Parasitology, School of Public HealthTehran University of Medical SciencesTehranIran
  4. 4.Department of Medical Physics, Faculty of Medical SciencesTarbiat Modares UniversityTehranIran
  5. 5.Nanosystems Research Team (NRTeam), Microbial Biotechnology and Biosafety DepartmentAgricultural Biotechnology Research Institute of Iran (ABRII)KarajIran
  6. 6.Research and Development Department of NanozinoTehranIran

Personalised recommendations