Development of antibody-based microarray assay for quantitative detection of aflatoxin B1

Authors

Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, University of Tehran, Tehran,

Abstract

BACKGROUND: Aflatoxin B1 (AFB1) is a toxic metabolite
produced by Aspergillus species that contaminates a wide range of
agricultural products. OBJECTIVES: This study was designed to
develop a rapid and highly sensitive immunoassay method in
microarray format for quantitative detection of AFB1 to evaluate
the potential of microarray platform for high-throughput screening,
which can be beneficial in food and feed industry. METHODS:
Following successful optimization, using an indirect competitive
immunoassay in dot blot format, AFB1-bovine serum albumin
(AFB1-BSA) conjugate was contact-printed onto 16 isolated subarrays
on multi-pad nitrocellulose coated slides; subsequently used
in competitive binding assays. RESULTS: Using the aforementioned
assay, AFB1 was determined from 15 pg/g to 3.04 ng/g working
range with detection limit (LOD) of 1 pg/g. To evaluate assay
performance in real food matrices, the authors spiked wheat
samples with different concentration of AFB1. After extraction,
working ranges of 0.11-4.15 ng/g with detection limit of 30pg/g was
determined. Good recoveries (94±9%) were obtained, demonstrating
accurate detection of AFB1 concentrations in wheat samples.
Assay procedure completed in 3 hours. CONCLUSIONS: The results
indicated that the proposed developed assay in microarray format
could be used for rapid and sensitive detection of AFB1in wheat
samples.

Keywords

Iranian Journal of Veterinary Medicine
Volume 8, Issue 2 - Serial Number 2
August 2014
Pages 135-142

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