Differentiation and quantification of the ochratoxin A producers Aspergillus ochraceus and Aspergillus westerdijkiae using PCR-DGGE
| Autor: | Mirna-Leonor Suàrez-Quiroz, Jean-Christophe Meile, Noël Durand, D. Montet, Sabine Schorr-Galindo, Angélique Fontana |
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| Rok vydání: | 2018 |
| Předmět: |
Ochratoxin A
Genes Fungal Biology Applied Microbiology and Biotechnology Polymerase Chain Reaction Sensitivity and Specificity 03 medical and health sciences chemistry.chemical_compound Électrophorèse sur gel Food science Ochratoxine Aspergillus westerdijkiae Mycotoxin DNA Fungal Pcr dgge 030304 developmental biology DNA Primers Aspergillus ochraceus 0303 health sciences 030306 microbiology Denaturing Gradient Gel Electrophoresis Microbiota Fungi General Medicine Sequence Analysis DNA Mycotoxins biology.organism_classification Ochratoxins stomatognathic diseases Aspergillus chemistry S50 - Santé humaine Q03 - Contamination et toxicologie alimentaires Primer (molecular biology) Sequence Alignment DNA Temperature gradient gel electrophoresis |
| Zdroj: | Journal of Basic Microbiology |
| ISSN: | 1521-4028 |
| Popis: | Ochratoxin A (OTA) is a nephrotoxic, teratogenic, immunotoxic, and carcinogenic mycotoxin which is produced in tropical zones mainly by Aspergillus carbonarius, A. niger, A. ochraceus, and A. westerdijkiae. A. ochraceus and A. westerdijkiae species are phenotypically and genomically very close but A. westerdijkiae produce OTA at a very higher level than A. ochraceus. These species have been differentiated recently. The DNA primer pairs which were drawn so far are not specific and a genomic region of the same size is amplified for both species or they are too specific, and in this case, the DNA of a single species is amplified. To help preventing OTA contamination of foodstuffs, the PCR‐DGGE (Denaturing Gradient Gel Electrophoresis) method was used to discriminate between A. ochraceus and A. westerdijkiae DNA fragments of the same size but with different sequences and thus faster access to a diagnosis of the toxigenic potential of the fungal microflora. The proposed methodology was able to differentiate A. westerdijkiae from A. ochraceus with only one primer pairs in a single run. A calibration based on initial DNA content was obtained from image analysis of the DGGE gels and a method of quantification of the two strains was proposed. |
| Databáze: | OpenAIRE |
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