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Applied and Environmental Microbiology, December 2002, p. 6273-6282, Vol. 68, No. 12
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.12.6273-6282.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Direct Identification in Food Samples of Listeria spp. and Listeria monocytogenes by Molecular Methods

Luca Cocolin,1* Kalliopi Rantsiou,1 Lucilla Iacumin,1 Carlo Cantoni,2 and Giuseppe Comi1

Dipartimento Scienze degli Alimenti, Università degli studi di Udine, 33100 Udine,1 Dipartimento di Scienze e Tecnologie Veterinarie per la Sicurezza degli Alimenti, Università degli studi di Milano, 20121 Milan, Italy2

Received 25 March 2002/ Accepted 17 September 2002

A new molecular approach for the detection and identification of Listeria spp. and Listeria monocytogenes in food is presented here. The method is based on the PCR amplification of a fragment of the iap gene from the five species belonging to the genus and on the analysis of the PCR products obtained by denaturing gradient gel electrophoresis (DGGE). The protocol was first optimized by using strains from international collections. Based on the differences present in the sequences amplified, it was possible to obtain species-specific DGGE migration that allowed fast and easy identification of L. monocytogenes, L. innocua, L. welshimeri, L. seeligeri, and L. ivanovii. Moreover, for L. monocytogenes serotypes, partial differentiation was possible. The optimized protocol was used for identification of Listeria strains traditionally isolated from food and for direct detection and identification of Listeria members in food after an overnight enrichment. Identification of 48 food isolates and direct detection of Listeria spp. in 73 food samples show the potential of the method that can be used as a fast screening test to investigate the presence of Listeria spp. and L. monocytogenes in food.

* Corresponding author. Mailing address: Food Science Department, Via Marangoni, 97, 33100 Udine, Italy. Phone: 39/0432/590-759. Fax: 39/0432/590-719. E-mail: luca.cocolin{at}dsa.uniud.it.

Applied and Environmental Microbiology, December 2002, p. 6273-6282, Vol. 68, No. 12
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.12.6273-6282.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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