1. Multiplex Detection of Aspergillus Species.
- Author
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Martínez-Culebras P, Selma MV, and Aznar R
- Subjects
- DNA, Fungal, Food Contamination, Food Microbiology, Vitis microbiology, Aspergillus classification, Aspergillus genetics, Multiplex Polymerase Chain Reaction, Real-Time Polymerase Chain Reaction
- Abstract
Multiplex real-time polymerase chain reaction (PCR) provides a fast and accurate DNA-based tool for the simultaneous amplification of more than one target sequence in a single reaction. Here a duplex real-time PCR assay is described for the simultaneous detection of Aspergillus carbonarius and members of the Aspergillus niger aggregate, which are the main responsible species for ochratoxin A (OTA) contamination in grapes. This single tube reaction targets the beta-ketosynthase and the acyl transferase domains of the polyketide synthase of A. carbonarius and the A. niger aggregate, respectively.Besides, a rapid and efficient fungi DNA extraction procedure is described suitable to be applied in wine grapes. It includes a pulsifier equipment to remove conidia from grapes which prevents releasing of PCR inhibitors.
- Published
- 2017
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