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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

Abstract

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.

Figure

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.

Keywords

Aflatoxin B1 Nanobiosensor FRET Immunoreaction Mycotoxin 

Notes

Acknowledgments

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

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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

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