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Applied and Environmental Microbiology, May 2009, p. 2925-2930, Vol. 75, No. 9
0099-2240/09/$08.00+0     doi:10.1128/AEM.02470-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Development and Application of a Novel Peptide Nucleic Acid Probe for the Specific Detection of Cronobacter Genomospecies (Enterobacter sakazakii) in Powdered Infant Formula{triangledown} ,{dagger}

C. Almeida,1,2 N. F. Azevedo,1,2 C. Iversen,3 S. Fanning,3 C. W. Keevil,2 and M. J. Vieira1*

IBB—Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal,1 Environmental Healthcare Unit, School of Biological Sciences, University of Southampton, Bassett Crescent East, SO16 7PX Southampton, United Kingdom,2 Centres for Food Safety & Food-borne Zoonomics, UCD Veterinary Sciences Centre, University College Dublin, Belfield, Dublin 4, Ireland3

Received 28 October 2008/ Accepted 23 February 2009

Here, we report a fluorescence in situ hybridization (FISH) method for rapid detection of Cronobacter strains in powdered infant formula (PIF) using a novel peptide nucleic acid (PNA) probe. Laboratory tests with several Enterobacteriaceae species showed that the specificity and sensitivity of the method were 100%. FISH using PNA could detect as few as 1 CFU per 10 g of Cronobacter in PIF after an 8-h enrichment step, even in a mixed population containing bacterial contaminants.

* Corresponding author. Mailing address: Centro de Engenharia Biólogica, Universidade do Minho, 4710-057 Braga, Portugal. Phone: 351 253 604411. Fax: 351 253 678986. E-mail: mjv{at}deb.uminho.pt

{triangledown} Published ahead of print on 6 March 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, May 2009, p. 2925-2930, Vol. 75, No. 9
0099-2240/09/$08.00+0     doi:10.1128/AEM.02470-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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