title: PCR method for the detection of potential ochratoxin-producing Aspergillus species in coffee beans.
abstract: Ochratoxin A (OA) is a nephrotoxic and carcinogenic mycotoxin that has been found in cereal and food commodities. Currently, Aspergillus carbonarius, A. niger and A. ochraceus have been recognized as the species responsible for OA in coffee beans. With the aim of developing multiplex-PCR for detection of these species in coffee bean samples, we first developed specific primers for A. niger detection. The primer designed (OPX7(372)) provided an amplicon of 372 pb in all A. niger stricto sensu isolates. The PCR assay developed for A. niger identification in pure culture was also successfully used for detecting this species in coffee beans. No cross-reaction was observed using DNA from coffee beans inoculated with closely related black aspergilli species. A multiplex-PCR method for detection of A. carbonarius, A. niger and A. ochraceus species in coffee beans was developed. From a single assay this method detected the amplicons of 809, 372, and 260 pb that represent the three most ochratoxigenic species found in coffee bean samples, i.e., A. carbonarius, A. niger and A. ochraceus, respectively.
authors.affiliation: Centro de Ciências Biológicas, Universidade Estadual de Londrina, P.O. Box 6001, 86051-990 Londrina-PR, Brazil.
authors.forename: Daniele
authors.initials: D
authors.lastname: Sartori
authors.forename: Márcia Cristina
authors.initials: MC
authors.lastname: Furlaneto
authors.forename: Mayra Kassawara
authors.initials: MK
authors.lastname: Martins
authors.forename: Maria Rosana
authors.initials: MR
authors.lastname: Ferreira de Paula
authors.forename: Aline Aparecida
authors.initials: AA
authors.lastname: Pizzirani-Kleiner
authors.forename: Marta Hiromi
authors.initials: MH
authors.lastname: Taniwaki
authors.forename: Maria Helena Pelegrinelli
authors.initials: MH
authors.lastname: Fungaro
mesh_terms: D001230:Aspergillus / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D040503:Coffea / Q000382:microbiology*; D017931:DNA Primers; D004271:DNA, Fungal / Q000235:genetics* / Q000302:isolation & purification; D016533:Mycological Typing Techniques; D009793:Ochratoxins / Q000096:biosynthesis*; D010802:Phylogeny; D016133:Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D009793:Ochratoxins