This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lambertz, S. T.
Right arrow Articles by Lindblad, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lambertz, S. T.
Right arrow Articles by Lindblad, M.
Agricola
Right arrow Articles by Lambertz, S. T.
Right arrow Articles by Lindblad, M.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, October 2008, p. 6060-6067, Vol. 74, No. 19
0099-2240/08/$08.00+0     doi:10.1128/AEM.00405-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Real-Time PCR Method for Detection of Pathogenic Yersinia enterocolitica in Food{triangledown}

S. Thisted Lambertz,1* C. Nilsson,1 S. Hallanvuo,2 and M. Lindblad1

Research and Development Department, National Food Administration, Uppsala, Sweden,1 Environmental and Food Research Laboratory (TavastLab), Municipal Joint Union for Public Health in Hämeenlinna Region, Hämeenlinna, Finland2

Received 18 February 2008/ Accepted 5 August 2008

The current methods for the detection of pathogenic Yersinia enterocolitica bacteria in food are time consuming and inefficient. Therefore, we have developed and evaluated in-house a TaqMan probe-based real-time PCR method for the detection of this pathogen. The complete method comprises overnight enrichment, DNA extraction, and real-time PCR amplification. Also included in the method is an internal amplification control. The selected primer-probe set was designed to use a 163-bp amplicon from the chromosomally located gene ail (attachment and invasion locus). The selectivity of the PCR method was tested with a diverse range (n = 152) of related and unrelated strains, and no false-negative or false-positive PCR results were obtained. The sensitivity of the PCR amplification was 85 fg purified genomic DNA, equivalent to 10 cells per PCR tube. Following the enrichment of 10 g of various food samples (milk, minced beef, cold-smoked sausage, fish, and carrots), the sensitivity ranged from 0.5 to 55 CFU Y. enterocolitica. Good precision, robustness, and efficiency of the PCR amplification were also established. In addition, the method was tested on naturally contaminated food; in all, 18 out of 125 samples were positive for the ail gene. Since no conventional culture method could be used as a reference method, the PCR products amplified from these samples were positively verified by using conventional PCR and sequencing of the amplicons. A rapid and specific real-time PCR method for the detection of pathogenic Y. enterocolitica bacteria in food, as presented here, provides a superior alternative to the currently available detection methods and makes it possible to identify the foods at risk for Y. enterocolitica contamination.

* Corresponding author. Mailing address: National Food Administration Research and Development Department, P.O. Box 622, SE-751 26 Uppsala, Sweden. Phone: 46 (0)18 17 55 62. Fax: 46 (0)18 17 14 94. E-mail: sula{at}slv.se

{triangledown} Published ahead of print on 15 August 2008.

Applied and Environmental Microbiology, October 2008, p. 6060-6067, Vol. 74, No. 19
0099-2240/08/$08.00+0     doi:10.1128/AEM.00405-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Lambertz, S. T., Nilsson, C., Hallanvuo, S. (2008). TaqMan-Based Real-Time PCR Method for Detection of Yersinia pseudotuberculosis in Food. Appl. Environ. Microbiol. 74: 6465-6469 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»