title: Multiplex Detection of Aspergillus Species.
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.
authors.affiliation: Department of Preventive Medicine, Public Health, Food Science and Technology, Bromatology, Toxicology, and Legal Medicine, University of Valencia, Valencia, Spain.|Department of Biotechnology, Institute of Agrochemistry and Food Technology (IATA-CSIC), Valencia, Spain.
authors.forename: Pedro
authors.initials: P
authors.lastname: Martínez-Culebras
authors.affiliation: Research Group on Quality Safety and Bioactivity of Plant Foods, Department of Food Science and Technology, CEBAS-CSIC, Murcia, Spain.
authors.forename: María Victoria
authors.initials: MV
authors.lastname: Selma
authors.affiliation: Department of Biotechnology, Institute of Agrochemistry and Food Technology (IATA-CSIC), Valencia, Spain. rosa.aznar@uv.es.|Department of Microbiology and Ecology and Spanish Type Culture Collection (CECT), University of Valencia, Valencia, Spain. rosa.aznar@uv.es.
authors.forename: Rosa
authors.initials: R
authors.lastname: Aznar
mesh_terms: D001230:Aspergillus / Q000145:classification* / Q000235:genetics*; D004271:DNA, Fungal; D005506:Food Contamination; D005516:Food Microbiology; D060885:Multiplex Polymerase Chain Reaction; D060888:Real-Time Polymerase Chain Reaction; D027843:Vitis / Q000382:microbiology
chemical_list: D004271:DNA, Fungal
keywords: Aspergillus carbonarius; Aspergillus niger aggregate; Grapes; Multiplex PCR; Mycotoxin; Ochratoxin A; Polyketide synthase; Real-time PCR; Wine