PCR-based detection and quantification of mycotoxin-producing fungi

Buslyk T.V., Rosalovsky V.P., Salyha Y.T.

SUMMARY. 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. This article describes the current state of problems regarding the contamination of agricultural products, caused by mycotoxins and mycotoxin-pro-ducing fungi, both in Ukraine and worldwide. The impacts of the exposure of animals to the most common mycotoxins, namely aflatoxins, fumonisins, ochratoxins, trichothecenes and zearalenone, are described. The prospects of using polymerase chain reaction (PCR) for the purpose of qualitative and quantitative detection of mycotoxin fungi and the ability to predict the production of mycotoxins by individual strains were analyzed. The estimated target regions in the genome of fungi can be potential markers for the identification of fungi species and determination of their chemotype. The focus is 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. We consider several PCR test systems for the detection of mycotoxinogenic molds, using sets of primers with structural and regulatory genes involved in the biosynthesis of aflatoxins (omt-1, nor1, ver1), fumonisins (fum1, fum13), trichocenes (tri1, tri3, tri5, tri6, tri12, tri13), zearalenone (PKS4, PKS13, ZEB1, ZEB2) and ochratoxin (pks, OTAnps). Considering the development of modern molecular genetic methods, the possibilities of using molecular methods are discussed, which include the analysis of DNA target regions for the differentiation of Fusarium, Aspergillus, Penicillium species.

Key words: contamination, mycotoxins, molds, genes, PCR

Tsitologiya i Genetika 2022, vol. 56, no. 1, pp. 22-38

• Institute of Animal Biology NAAS, V. Stus Street 38, Lviv 79034, Ukraine

E-mail: yursalyha yahoo.com

Buslyk T.V., Rosalovsky V.P., Salyha Y.T. PCR-based detection and quantification of mycotoxin-producing fungi, Tsitol Genet., 2022, vol. 56, no. 1, pp. 22-38.