PCR Assay for Identification of Aspergillus Carbonarius and Aspergillus Japonicus

Black Aspergilli, and in particular Aspergillus carbonarius, are the main causes of contamination of grapes and their by-products by ochratoxin A. A PCR-based method was developed to detect DNA of A. carbonarius and A. japonicus. Two pairs of primers (CARBO1/2 and JAPO1/2) designed from the calmodulin gene, produced PCR products of 371 and 583 bp for A. carbonarius and A. japonicus, respectively. Primer specificity was tested with DNA of 107 strains belonging to Aspergillus section Nigri isolated mostly from grapes in Europe. The sensitivity of primers CARBO1/2 and JAPO1/2 was 12.5 pg when using pure total genomic DNA of the two species. The developed primers provide a powerful tool for detection of the main ochratoxigenic producing Aspergillus species in grapes.