Current PCR-based methods for the detection of mycotoxigenic fungi in complex food and feed matrices

Abstract

Mycotoxins are toxic secondary fungal metabolites produced by certain types of filamentous fungi, such as Aspergillus, Fusarium, and Penicillium spp. Mycotoxigenic fungi and their produced mycotoxins are considered to be an important issue in food and feed safety due to their toxic effects like carcinogenicity, immunosuppression, neurotoxicity, nephrotoxicity, and hepatotoxicity on humans and animals. To boost the safety level of food and feedstuff, detection and identification of toxins are essential at critical control points across food and feed chains. Zero-tolerance policies by the European Union and other organizations about the extreme low level of tolerance of mycotoxins contamination in food and feed matrices have led to an increasing interest to design more sensitive, specific, rapid, cost-effective, and safer to use mycotoxigenic fungi detection technologies. Hence, many mycotoxigenic fungi detection technologies have been applied to measure and control toxins contamination in food and feed substrates. PCR-based mycotoxigenic fungi detection technologies, such as conventional PCR, real-time PCR, nested PCR, reverse transcriptase (RT)-PCR, loop-mediated isothermal amplification (LAMP), in situ PCR, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR DGGE), co-operational PCR, multiplex PCR, DNA arrays, magnetic capture-hybridization (MCH)-PCR and restriction fragment length polymorphism (RFLP), would contribute to our understanding about different mycotoxigenic fungi detection approaches and will enhance our capability about mycotoxigenic fungi identification, isolation and characterization at critical control points across food and feed chains. We have assessed the principles, results, the limit of detection, and application of these PCR-based detection technologies to alleviate mycotoxins contamination problem in complex food and feed substrates. The potential application of these detection technologies can reduce mycotoxins in complex food and feed matrices.

Applicability of Smart Tools in Vegetable Disease Diagnostics

“…With regard to plant pathogen detection/quantification, we found many protocols for vegetable phytopathogen species, but true validation reports, as required by the Minimal Performance Criteria (MIQE) [36], are not available. The in-lab verification followed by the full validation of published and forthcoming methods is a necessity for researchers to rely on their performance after the provision of comparable results across different laboratories [36][37][38][39][40]. To ensure uniform test performance across laboratories, international standards and guidelines have been issued by the International Organization for Standardization (ISO) that are used by accredited testing laboratories [41].…”

Fungal Contaminants and Mycotoxins in Nuts

“…After this step, nucleic acids can be purified to eliminate impurities [102]. Species-specific PCR methodologies targeting conserved genes or regions of taxonomical interest or by focusing on the mycotoxigenic genes have been extensively applied for identification of mycotoxigenic fungal contaminants [103][104][105][106].…”

Patulin-producing mold, toxicological, biosynthesis, and molecular detection of patulin

“…The most critical factor in PCR-based detection is primer design which should meet the criteria like GC content (40-60%), Tm values (50-80ºC), 5'-end stability, 3'-end specificity, and minimum interaction of primer-primer and hairpins. Detection of PCR amplification on conventional PCR uses agarose gels at the final stage of the PCR reaction (Ur Rahman et al, 2020). Real-time PCR allows the detection of PCR amplification during the early stage of the reaction and detects and quantifies DNA of toxigenic mold in food product.…”