Issue 19, 2018

Development of a FRET-based fluorescence aptasensor for the detection of aflatoxin B1 in contaminated food grain samples

Abstract

The present study aimed to develop an aptamer-based FRET detection strategy for the specific and sensitive detection of AFB1 in contaminated food grains. The study comprises generation of ssDNA aptamers against AFB1 by whole-cell SELEX and their application in a FRET-based platform utilizing graphene oxide (GO) and quantum dots (QDs). The generated aptamers were characterized to determine their specificity and sensitivity using indirect ELISA where AFB1–OVA was used as a coating antigen. Among the aptamers generated, the ATB1 aptamer showed good reactivity and selectivity against AFB1. This aptamer was further characterized to determine its secondary structure and KD value, which was found to be 5.9 kcal mol−1. The characterized aptamers were conjugated onto Cd/Se quantum dots to develop a fluorimetric system for the detection of aflatoxin B1 using a graphene oxide platform. The presence of graphene oxide quenches the fluorescence ability of the quantum dots due to π–π stacking interactions between the aptamer and GO. Upon target addition, the aptamer forms a complex with aflatoxin B1 thereby restoring the fluorescence intensity. The developed assay shows a linear response from 0.002 μg μl−1 to 0.2 μg μl−1 with a detection limit of 0.004 μg μl−1 for the AFB1 standard toxin and showed no cross-reactivity with other closely related mycotoxins. To validate the reliability of the developed method, several field samples spiked with AFB1 were included in this study and the results obtained were cross verified using a standard commercial AFB1 kit. In conclusion, the developed method may find good utility in routine food testing laboratories for risk assessment of AFB1.

Graphical abstract: Development of a FRET-based fluorescence aptasensor for the detection of aflatoxin B1 in contaminated food grain samples

Article information

Article type
Paper
Submitted
11 1 2018
Accepted
15 2 2018
First published
14 3 2018
This article is Open Access
Creative Commons BY license

RSC Adv., 2018,8, 10465-10473

Development of a FRET-based fluorescence aptasensor for the detection of aflatoxin B1 in contaminated food grain samples

A. Kumar Y. V. V., R. R. M., A. J., V. Mudili and S. Poda, RSC Adv., 2018, 8, 10465 DOI: 10.1039/C8RA00317C

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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