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

Abstract

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