Multiplex Detection of Aspergillus Species (real-time PCR)

Detail:

Series: Methods in Molecular Biology | Volume: 1542 | Pub. Date: Dec-11-2016 | Page Range: 261-268 | DOI: 10.1007/978-1-4939-6707-0_17

Year prepared: 2016

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.

Laboratory Protocols

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