HIGHLY SENSITIVE FRET-BASED FLUORESCENCE IMMUNOASSAY FOR DETECTING OF AFLATOXIN B1 USING MAGNETIC/SILICA CORE-SHELL AS A SIGNAL INTENSIFIER

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KALARESTAGHI ALIREZA; BAYAT MANSOUR; HASHEMI SEYED JAMAL; RAZAVILAR VADOOD

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Journal: IRANIAN JOURNAL OF BIOTECHNOLOGY Year:2015 | Volume:13 | Issue:3 Page(s): 25-31

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Title

HIGHLY SENSITIVE FRET-BASED FLUORESCENCE IMMUNOASSAY FOR DETECTING OF AFLATOXIN B1 USING MAGNETIC/SILICA CORE-SHELL AS A SIGNAL INTENSIFIER

Author(s)

KALARESTAGHI ALIREZA | BAYAT MANSOUR | HASHEMI SEYED JAMAL | RAZAVILAR VADOOD

Keywords

AFLATOXIN B1

FRET

MAGNETIC/SILICA CORE SHELL

NANOBIOSENSOR

Abstract

Background: Recently, some new nanobiosensors using different nanoparticles or microarray systems for detection ofmycotoxins have been designed. However, rapid, sensitive and early detection of aflatoxicosis would be very helpful todistinguish high-risk persons.Objectives: We report a highly sensitive competitive immunoassay using magnetic/silica core shell as a signal intensifierfor the determination of aflatoxin B1 using fluorescence resonance energy transfer (FRET) from Cd/Te quantum dots (antiaflatoxinB1 antibody immobilized on the surface of Cd/Te quantum dots) to Rhodamine 123 (Rho 123-labeled aflatoxinB1 bound to albumin). The specific immune-reaction between the anti-aflatoxin B1 antibody on the QDs and the labeledaflatoxinB1 brings the Rho 123 fluorophore (acting as the acceptor) and the QDs (acting as the donor) in close spatialproximity and causes FRET to occur upon photo-excitation of the QDs. Using magnetic/silica core shell to intensify theobtained signal is the novelty of this study.Materials and Methods: Cd/Te QDs were synthesized by the simultaneous reduction of cadmium chloride and telluriumin the presence of sodium borohydride under nitrogen atmosphere. Magnetic nanoparticles were synthesized using FeSO4 and FeCl3 (1: 2 molar ratio) and ammonia as an oxidizing agent under nitrogen atmosphere. The prepared magneticnanoparticles shelled by silica using tetraethoxysilane in the presence of ammonia. Nanoparticles synthesis and monodispersity confirmed by TEM. Immobilization of Cd/Te QDs to antibodies and labeling of aflatoxin B1-albumin by Rho 123 were performed by EDC/NHS reaction in reaction mixture buffer, pH 6, at room temperature.Results: By using the magnetic/silica core shell sensitivity of the system changed from 2×10-11 in our previous study to2×10-12 in this work. The feasibility of the method established by the detection of aflatoxin B1 in spiked human serum.There is a linear relationship between the decreased fluorescence intensity of Rho 123 with increasing concentration ofaflatoxin B1 in spiked samples, over the range of 0.01-0.06 μmol.mL-1.Conclusions: This homogeneous competitive detection scheme is simple, rapid and efficient, and does not require multipleseparation steps and excessive washing.

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APA: Copy

KALARESTAGHI, ALIREZA, BAYAT, MANSOUR, HASHEMI, SEYED JAMAL, & RAZAVILAR, VADOOD. (2015). HIGHLY SENSITIVE FRET-BASED FLUORESCENCE IMMUNOASSAY FOR DETECTING OF AFLATOXIN B1 USING MAGNETIC/SILICA CORE-SHELL AS A SIGNAL INTENSIFIER. IRANIAN JOURNAL OF BIOTECHNOLOGY, 13(3), 25-31. SID. https://sid.ir/paper/693909/en

Vancouver: Copy

KALARESTAGHI ALIREZA, BAYAT MANSOUR, HASHEMI SEYED JAMAL, RAZAVILAR VADOOD. HIGHLY SENSITIVE FRET-BASED FLUORESCENCE IMMUNOASSAY FOR DETECTING OF AFLATOXIN B1 USING MAGNETIC/SILICA CORE-SHELL AS A SIGNAL INTENSIFIER. IRANIAN JOURNAL OF BIOTECHNOLOGY[Internet]. 2015;13(3):25-31. Available from: https://sid.ir/paper/693909/en

IEEE: Copy

ALIREZA KALARESTAGHI, MANSOUR BAYAT, SEYED JAMAL HASHEMI, and VADOOD RAZAVILAR, “HIGHLY SENSITIVE FRET-BASED FLUORESCENCE IMMUNOASSAY FOR DETECTING OF AFLATOXIN B1 USING MAGNETIC/SILICA CORE-SHELL AS A SIGNAL INTENSIFIER,” IRANIAN JOURNAL OF BIOTECHNOLOGY, vol. 13, no. 3, pp. 25–31, 2015, [Online]. Available: https://sid.ir/paper/693909/en

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