Abstract
Aflatoxin contamination in agricultural products has posed serious health hazards and brought huge economic loss in the food and feed industries. Monitoring aflatoxins in various foods and feeds has become a crucial means to protect public health. This study aimed to report an immuno-loop-mediated isothermal amplification (iLAMP) assay by using an anti-idiotypic nanobody-phage for on-site and rapid detection of aflatoxin in real samples. The iLAMP method was developed on the basis of a competitive immunoassay and LAMP reaction performed in a simple water bath. This method can provide visualized test results: violet color represents positive samples while sky blue represents negative. The visual detection limits of iLAMP for aflatoxin B1, B2, G1, and G2 in peanut samples were 1.6, 1.6, 3.2, and 16 μg/kg, respectively. The developed assay was verified with high performance liquid chromatography (HPLC) for the analysis of aflatoxins in peanuts, which demonstrated that the iLAMP method can be applied to the detection of aflatoxin in real samples. The novel iLAMP assay eliminates the need for aflatoxin conjugates, the antibody labeling process, and special equipment, and offers an alternative to existing methods with advantages of time-saving, cost-effectiveness, and ease-of-use.
Highlights
Aflatoxins are highly toxic secondary metabolites produced by fungal species of Aspergilli, especially Aspergillus flavus and Aspergillus parasiticus [1]
We developed an immuno-Loop-mediated isothermal amplification (LAMP) assay for aflatoxin detection based on anti-aflatoxin monoclonal antibody 1C11 and anti-idiotypic nanobody-phage V2–5 specific for mAb 1C11
The isothermal amplification (iLAMP) method was developed on the basis of a competitive immunoassay and LAMP reaction
Summary
Aflatoxins are highly toxic secondary metabolites produced by fungal species of Aspergilli, especially Aspergillus flavus and Aspergillus parasiticus [1]. They are listed as group I liver carcinogens by the International Agency for Research on Cancer (IARC) [2]. Aflatoxin contamination poses a serious threat to human health as well as to the food industry, and has received considerable public attention over the past decades [5]. Monitoring aflatoxins in various food and feeds has become a crucial means to protect public health. Many methods have been developed for aflatoxin detection to meet public concerns about food safety and increasingly stringent analytical requirements, ranging from confirmatory tests in official laboratories to rapid on-site tests in the field [6]
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