Evaluation of a multiplex PCR assay for concurrent detection of four major mycotoxigenic fungi from foods
| Autor: | R. Rashmi, H.S. Murali, Siva R. Uppalapati, Harsh Vardhan Batra, R. Shylaja, Mudili Venkata Ramana |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Ochratoxin A
Fusarium Aflatoxin Trichothecene Food Contamination Fumonisins Sensitivity and Specificity Applied Microbiology and Biotechnology Microbiology chemistry.chemical_compound Fumonisin Multiplex polymerase chain reaction High performance thin layer chromatography Food science DNA Fungal Mycotoxin Chromatography High Pressure Liquid DNA Primers biology Penicillium food and beverages General Medicine Mycotoxins biology.organism_classification Ochratoxins Aspergillus chemistry Food Microbiology Trichothecenes Multiplex Polymerase Chain Reaction Biotechnology |
| Zdroj: | Journal of Applied Microbiology. 114:819-827 |
| ISSN: | 1364-5072 |
| DOI: | 10.1111/jam.12100 |
| Popis: | Aim To develop and evaluate a multiplex polymerase chain reaction assay (mPCR) for the concurrent detection of four major mycotoxin metabolic pathway genes, viz. nor1 (aflatoxin), Tri6 (trichothecene), FUM13 (fumonisin) and otanps (ochratoxin A). Methods and Results A mPCR assay with competitive internal amplification control, employing specific primers for each of the aforementioned four genes, was optimized and validated using 10 reference strains and 60 pure culture isolates. The standardized mPCR assay detected all four mycotoxin metabolic genes in artificially contaminated maize samples with a sensitivity of 2 × 103 CFU g−1 for nor1-positive Aspergillus strains, Tri6 and FUM13-positive Fusarium strains and 2 × 104 CFU g−1 for otanps-positive Penicillium strains. When the developed mPCR assay was applied to 40 natural foods, 35% (14 of 40) of the samples were contaminated with either one or more mycotoxins. The mPCR results were further evaluated with high-performance liquid chromatography (HPLC), and in general, both the methods provided unequivocal results. Conclusion The current mPCR assay is a rapid and reliable tool for simultaneous specific and sensitive detection of aflatoxigenic Aspergillus strains, trichothecene- and fumonisin-producing Fusarium strains, and ochratoxigenic Penicillium species from naturally contaminated foods. Significance and Impact of the Study This mPCR assay could be a supplementary strategy to current conventional mycotoxin analytical techniques such as thin-layer chromatography (TLC), high performance thin layer chromatography, HPLC, etc., and a reliable tool for high-throughput monitoring of major mycotoxin-producing fungi during the processing steps of food and feed commodities. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text