PCR detection assays for the ochratoxin-producing Aspergillus carbonarius and Aspergillus ochraceus species Bele ´n Patin ˜o a , Amaia Gonza ´lez-Salgado b ,M a Teresa Gonza ´lez-Jae ´n b , Covadonga Va ´zquez a, * a Departamento de Microbiologı ´a III, Universidad Complutense de Madrid, Spain b Departamento de Gene ´tica, Universidad Complutense de Madrid, Spain Received 6 April 2004; received in revised form 3 November 2004; accepted 5 February 2005 Abstract Two PCR assays have been developed to detect Aspergillus carbonarius and Aspergillus ochraceus , considered the main sources of ochratoxin A (OTA) contaminating commodities, particularly grapes, coffee and derivatives, in warm climates. The species specific primers have been designed on the basis of ITS (internal transcribed spacers of rDNA units) sequence comparisons obtained from Aspergillus strains and have been tested in a number of strains from different origins and hosts. These PCR assays, based on multi-copy sequences, are highly sensitive and specific and represent a good tool for an early detection of OTA-producing Aspergillus species and to prevent OTA entering the food chain. D 2005 Elsevier B.V. All rights reserved. Keywords: Aspergillus carbonarius ; Aspergillus ochraceus ; Ochratoxin A; PCR; Detection; ITS 1. Introduction Ochratoxin A (OTA) is a secondary metabolite produced by Aspergillus and Penicillium species. This mycotoxin has been shown to have nephrotoxic, inmunotoxic, genotoxic and teratogenic properties to- wards several animal species, and has been classified by International Agency for Research on Cancer as possible carcinogen to humans (group 2B) (IARC, 1993). OTA occurs in various foodstuffs and beve- rages including a variety of cereals, beans, ground- nuts, spices, dried fruits, coffee, milk, wine and beer (Varga et al., 2001; Caban ˜es et al., 2002; Petzinger and Weidenbach, 2002; Serra et al., 2003), and its maximum limit on several commodities for human consumption are under legal regulation. Two Aspergillus sections are known to produce OTA: the section Circumdati (also called the Asper- gillus ochraceus group) and the section Nigri (Asper- gillus carbonarius and Aspergillus niger )(Te ´ren et 0168-1605/$ - see front matter D 2005 Elsevier B.V. All rights reserved. doi:10.1016/j.ijfoodmicro.2005.02.011 * Corresponding author. Department of Microbiology III, Faculty of Biology, University Complutense of Madrid, Jose ´ Antonio Nova ´is 2, 28040-Madrid, Spain. Tel.: +34 913 944 969; fax: +34 913 944 964. E-mail address: covi@bio.ucm.es (C. Va ´zquez). International Journal of Food Microbiology 104 (2005) 207 – 214 www.elsevier.com/locate/ijfoodmicro