Abstract
The protocol for detecting Aspergillus flavus was based on a simple polymerase chain reaction with high sensitivity and specificity. The specific primers were designed relying on the aflatoxin biosynthesis gene clusters. The limit of detection (LOD) is 2 ng/assay for single DNA and 3.75 ng/assay for mixed DNA. The direct extraction of DNA from the infected samples is used for the protocol. The LOD of the protocol in the detection of A. flavus is 40 ng of total DNA. The results also indicated that the sensitivity and specificity of the optimised protocol was twenty times higher than that of the conventional morphological method. This protocol could be used for monitoring A. flavus in stored food or feed, especially in the detection of a potential aflatoxigenic risk in stored peanut and corn kernels, using a rapid, effective and high-throughput method, even in the case of low-level infections.
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