title: Identification of ochratoxin A producing Aspergillus carbonarius and A. niger clade isolated from grapes using the loop-mediated isothermal amplification (LAMP) reaction.
abstract: OBJECTIVE
  To develop two assays based on the loop-mediated isothermal amplification (LAMP) of DNA for the quick and specific identification of Aspergillus carbonarius and ochratoxigenic strains of the Aspergillus niger clade isolated from grapes.
  
  
  RESULTS
  Two sets of primers were designed based on the polyketide synthase genes involved or putatively involved in ochratoxin A (OTA) biosynthesis in A. carbonarius and A. niger clade. Hydroxynaphthol blue was used as indirect method to indicate DNA amplification. The limit of detection of both assays was comparable to that of a PCR reaction. Specificities of the reactions were tested using DNA from different black aspergilli isolated from grapes. The two LAMP assays were then used to identify A. carbonarius and ochratoxigenic A. niger and A. awamori grown in pure cultures without a prior DNA extraction.
  
  
  CONCLUSIONS
  The two LAMP assays permitted to quickly and specifically identify DNA from OTA-producing black aspergilli, as well as isolates grown in pure culture.
  
  
  CONCLUSIONS
  Monitoring vineyards for the presence of OTA-producing strains is part of the measures to minimize the occurrence of OTA in grape products. The two LAMP assays developed here could be potentially used to speed the screening process of vineyards for the presence of OTA-producing black aspergilli.
authors: Plant Pathology Group, Institute of Integrative Biology, ETH Zurich, Zürich, Switzerland.
authors: M
authors: M
authors: Storari
authors: R
authors: R
authors: von Rohr
authors: I
authors: I
authors: Pertot
authors: C
authors: C
authors: Gessler
authors: G A L
authors: GA
authors: Broggini
mesh_terms: D001230:Aspergillus / Q000145:classification* / Q000302:isolation & purification / Q000378:metabolism; D001234:Aspergillus niger / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D005506:Food Contamination; D005516:Food Microbiology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009793:Ochratoxins / Q000096:biosynthesis*; D048630:Polyketide Synthases / Q000235:genetics; D012680:Sensitivity and Specificity; D027843:Vitis / Q000382:microbiology*
chemical_list: D017931:DNA Primers; D009793:Ochratoxins; C025589:ochratoxin A; D048630:Polyketide Synthases