PCR-Based Detection and Quantification of Mycotoxin-Producing Fungi

One of the main factors reducing the quality of agricultural products can be contamination with molds capable of producing mycotoxins that pose a threat to both human and animal health. In modern conditions of closer attention to feed safety, the issues of contamination with mycotoxins and mycotoxigenic fungi are especially relevant. The review describes the current state of the problem regarding contamination of agricultural products with mycotoxins and mycotoxigenic molds in Ukraine and worldwide. The impacts of exposure of animals to the most common mycotoxins, such as aflatoxins, fumonisins, ochratoxins, trichothecenes, and zearalenone, are described in the article. The prospects of using polymerase chain reaction (PCR) for the qualitative and quantitative detection of mycotoxin-producing fungi, as well as for a possibility of predicting the production of mycotoxins by individual strains, were analyzed. The target regions in the fungi genome were assessed as potential markers for the identification of fungal species and their chemotype differentiation. The attention was focused on the relatively simple PCR method with electrophoretic detection of amplification products and real-time PCR (RT-PCR) using SYBR Green dye interface technologies and TaqMan fluorescent oligonucleotide probes. Several PCR test systems for the detection of mycotoxinogenic molds are considered using sets of primers oriented to the structural and regulatory genes involved in the biosynthesis of aflatoxins (omt-1, nor1, and ver1), fumonisins (fum1 and fum13), trichothecenes (tri1, tri3, tri5, tri6, tri12, and tri13), zearalenone (PKS4, PKS13, ZEB1, and ZEB2), and ochratoxin (pks and OTAnps). Considering the progress of modern molecular genetic methods, the possibilities of using molecular methods are discussed, which include the analysis of DNA target regions for differentiation of Fusarium, Aspergillus, and Penicillium species.