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Applied and Environmental Microbiology, October 1999, p. 4688-4692, Vol. 65, No. 10
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Detection and Differentiation of Listeria spp. by a Single Reaction Based on Multiplex PCR

Andreas Bubert,1,2,* Inge Hein,3 Marcus Rauch,1 Angelika Lehner,3 ByoungSu Yoon,4 Werner Goebel,1 and Martin Wagner3

Lehrstuhl für Mikrobiologie, Theodor-Boveri-Institut für Biowissenschaften, Universität Würzburg, 97074 Würzburg,1 and Microbiological Analytics, Merck KGaA, 64271 Darmstadt,2 Germany; Institut für Milchhygiene und Milchtechnologie, Veterinärmedizinische Universität Wien, Vienna, Austria3; and Department of Biology, Kyonggi University, Suwon, Kyonggi-Do 442-760, Korea4

Received 8 March 1999/Accepted 23 June 1999

The iap gene encodes the protein p60, which is common to all Listeria species. A previous comparison of the DNA sequences indicated conserved and species-specific gene portions. Based on these comparisons, a combination consisting of only five different primers that allows the specific detection and differentiation of Listeria species with a single multiplex PCR and subsequent gel analysis was selected. One primer was derived from the conserved 3' end and is specific for all Listeria species; the other four primers are specific for Listeria monocytogenes, L. innocua, L. grayi, or the three grouped species L. ivanovii, L. seeligeri, and L. welshimeri, respectively. The PCR method, which also enables the simultaneous detection of L. monocytogenes and L. innocua, was evaluated against conventional biotyping with 200 food hygiene-relevant Listeria strains. The results indicated the superiority of this technique. Thus, this novel type of multiplex PCR may be useful for rapid Listeria species confirmation and for identification of Listeria species for strains isolated from different sources.

* Corresponding author. Mailing address: Merck KGaA, Scientific Laboratory Products, Microbiological Analytics, Frankfurter Strasse 250, 64293 Darmstadt, Germany. Phone: 49-6151-727661. Fax: 49-6151-726904. E-mail: andreas.bubert{at}merck.de.

Applied and Environmental Microbiology, October 1999, p. 4688-4692, Vol. 65, No. 10
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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