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Reliability of Listeria monocytogenes Identification by Specific PCR Assessed by Phenotypic and Genotypic Techniques

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Abstract

This study evaluates the possibility of using polymerase chain reaction (PCR) for rapid identification of food-borne Listeria monocytogenes as an alternative to API Listeria system and estimates the incidence of API Listeria misidentifications in food-borne Listeria species. A total of 198 strains, 11 L. monocytogenes, 28 other Listeria species, and 159 food isolates were phenotypically and genotypically characterized by API Listeria profiles and randomly amplified polymorphic DNA (RAPD) profiles, respectively. They were also tested for PCR amplification using genus- and species-specific primers. Clustering analysis of phenotypic and genotypic data showed discrepancies in species identification of some isolates by API Listeria profiles. Their identities were confirmed by 16S rDNA sequencing, and thus, it was revealed that 33% of Listeria innocua and 19% of Listeria welshimeri were misidentified as L. monocytogenes by API Listeria profiles. Reliable identification of L. monocytogenes was obtained by LM1–LM2 specific primers which allowed PCR amplification only in reference strains and isolates previously identified as L. monocytogenes by RAPD and 16S rDNA sequence analysis. These results corroborate the suitability of specific PCR as a rapid and accurate test for the identification of L. monocytogenes, avoiding misidentification with other Listeria species commonly found in food products.

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Abbreviations

RAPD:

randomly amplified polymorphic DNA

CECT:

Spanish type culture collection, Spain

References

  • Allerberger F, Dierich M, Petranyi G, Lalic M, Bubert A (1997) Non-haemolytic strains of Listeria monocytogenes detected in milk products using VIDAS Immunoassay Kit. Zent Hyg Umweltmed 200:189–195

    CAS  Google Scholar 

  • Anon (1997) Microbiology of food and animal feeding stuffs—horizontal method for the detection and enumeration of Listeria monocytogenes—Part 1: Detection, EN ISO 11290-1. International Organisation for Standardisation, Geneva

    Google Scholar 

  • Anon (1999) Opinion of the Scientific Committee on Veterinary Measures Relating to Public Health on Listeria monocytogenes. Brussels, Belgium: European Commission. Health and Consumer Protection Directorate-General

  • Anon (2005a) Commision regulation No. 2073/2005 on microbiological criteria for food stuffs. J Eur Union L388:1–26

    Google Scholar 

  • Anon (2005b) Microbiology of food and animal feeding stuffs—horizontal method for the detection and enumeration of Listeria monocytogenes—Part 1: Detection. Amendment 1: Modification of the isolation media and the haemolysis test, and inclusion of precision data (ISO 11290-1:1996/AM1:2004), (2005). EN ISO 11290-1:1997/A1:2005. International Organisation for Standardisation, Geneva

    Google Scholar 

  • Aznar R, Alarcón B (2002) On the specificity of PCR detection of Listeria monocytogenes in food: a comparison of published primers. Syst Appl Microbiol 25:109–119

    Article  CAS  Google Scholar 

  • Bille J, Catimel B, Bannerman E, Jacquet C, Yersin MN, Caniaux I, Monget D, Rocourt J (1992) API Listeria, a new and promising one-day system to identify Listeria isolates. Appl Environ Microbiol 58:1857–1860

    CAS  Google Scholar 

  • Border PM, Howard JJ, Plastow GS, Siggens KW (1990) Detection of Listeria species and Listeria monocytogenes using polymerase chain reaction. Lett Appl Microbiol 11:158–162

    Article  CAS  Google Scholar 

  • Brosius J, Dull TJ, Sleeter DD, Noller HF (1981) Gene organization and primary structure of a ribosomal RNA operon from Escherichia coli. J Mol Biol 148(2):107–27

    Article  CAS  Google Scholar 

  • Clark AG, McLaughlin J (1997) A simple colour test based on an alanyl peptidase reaction which differentiates Listeria monocytogenes from other Listeria species. J Clin Microbiol 35:2155–2156

    CAS  Google Scholar 

  • Cummings AJ, Fielding AK, McLaughlin J (1994) Listeria ivanovii infection in a patient with AIDS. J Infect 28:89–91

    Article  Google Scholar 

  • Furrer B, Candrian U, Hoefelein C, Luethy J (1991) Detection and identification of Listeria monocytogenes in cooked sausage products and in milk by in vitro amplification of haemolysin gene fragments. J Appl Bacteriol 70:372–379

    CAS  Google Scholar 

  • Gouws PA, Liedemann I (2005) Evaluation of diagnostic PCR for the detection of Listeria monocytogenes in food products. Food Tech Biotech 43:201–205

    CAS  Google Scholar 

  • Greenwood M, Willis C, Doswell P, Allen G, Pathak K (2005) Evaluation of chromogenic media for the detection of Listeria species in food. J Appl Microbiol 99:1340–1345

    Article  CAS  Google Scholar 

  • Hitchins AD (1995) Listeria monocytogenes. In: Food and Drug Administration (ed) Bacteriological analytical manual. 8th ed. Revision A. AOAC International, Gaithersburg, MD, USA, pp. 10.01–10.13

  • Johnson J, Jinneman K, Stelma G, Smith BG, Lye D, Messer J, Ulaszek J, Evsen L, Gendel S, Bennett RW, Swaminathan B, Pruckler J, Steigerwalt A, Kathariou S, Yildirim S, Volokhov D, Rasooly A, Chizhikov V, Wiedmann M, Fortes E, Duvall RE, Hitchins AD (2004) Natural atypical Listeria innocua strains with Listeria monocytogenes pathogenicity island 1 genes. Appl Environ Microbiol 70:4256–4266

    Article  CAS  Google Scholar 

  • Karpísková R, Pejchalová M, Mokrosová J, Vytrasová J, Smuharová P, Ruprich J (2000) Application of a chromogenic medium and the PCR method for the rapid confirmation of L. monocytogenes in foodstuffs. J Microbiol Meth 41:267–271

    Article  Google Scholar 

  • Ludwig W, Strunk O, Westram R et al (2004) ARB: a software environment for sequence data. Nucl Acids Res 32:1363–1371

    Article  CAS  Google Scholar 

  • Malorny B, Tassios PT, Radström P, Cook N, Wagner M, Hoorfar J (2003) Standardization of diagnostic PCR for the detection of foodborne pathogens. Int J Food Microbiol 83:39–48

    Article  CAS  Google Scholar 

  • McLauchlin J (1997) The identification of Listeria species. Int J Food Microbiol 38:77–81

    Article  CAS  Google Scholar 

  • Pinto M, Burri S, Mena C, Almeida G, Carneiro L, Teixeira P, Gibbs PA (2001) Comparison of oxford agar, PALCAM and Listeria monocytogenes blood agar for the recovery of L. monocytogenes from foods and environmental samples. Food Control 12:512–514

    Article  Google Scholar 

  • Pitcher DG, Saunders NA, Owen RJ (1989) Rapid extraction of bacterial genomic DNA with guanidium thiocynate. Lett Appl Microbiol 8:151–156

    Article  CAS  Google Scholar 

  • Rijpens N, Vlaemynck G, Rossau R, Herman L, Jannes G (1998) Unidentified Listeria-like bacteria isolated from cheese. Lett Appl Microbiol 27:198–202

    Article  CAS  Google Scholar 

  • Rocourt J (1999) The genus Listeria and Listeria monocytogenes: phylogenetic position, taxonomy, and identification. In: Ryser ET, Marth EH (eds) Listeria, listeriosis, and food safety, 2nd ed. Marcel Dekker, New York, NY, USA, pp 1–20

    Google Scholar 

  • Rocourt J, Hof H, Schrettenbrunner A, Malinverni R, Bille J (1986) Acute purulent Listeria seeligeri in an immunocompetent adult. Schweiz Med Wochenschr 116:248–251

    CAS  Google Scholar 

  • Seeliger H, Jones D (1986) Genus Listeria. In: Williams and Wilkins (ed) Bergey’s manual of systematic bacteriology. Vol. II. Baltimore, USA, pp: 1235–1245

  • Sneath PHA, Sokal RR (1973) Numerical taxonomy. Freeman, San Francisco

    Google Scholar 

  • Vaneechoutte M, Boerlin P, Tichy HV, Bannerman E, Jager B, Bille J (1998) Comparison of PCR-based DNA fingerprinting techniques for the identification of Listeria species and their use for atypical Listeria isolates. Int J Syst Bacteriol 1:127–139

    Article  Google Scholar 

  • Vlaemynck G, Lafarge V, Scotter S (2000) Improvement of the detection of Listeria monocytogenes by the application of ALOA, a diagnostic, chromogenic isolation medium. J Appl Microbiol 88:430–441

    Article  CAS  Google Scholar 

  • Walker JK, Morgan JH, McLauchin J, Grant KA, Shallcross JA (1994) Listeria innocua isolated from a case of ovine meningoencephalitis. Vet Microbiol 42:245–253

    Article  CAS  Google Scholar 

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Acknowledgements

This work has been supported by “Comisión Interministerial de Ciencia y Tecnología” (CICYT) grant 1FD97-0549. We thank Beatriz Pinto for technical assistance, IPROMA Laboratory (Castellón) for the collaborative work and the CECT for providing reference strains. English text revised by F. Barraclough.

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Authors and Affiliations

  1. Departamento de Microbiología y Ecología, Universitat de València, València, Spain

    Rosa Aznar & Patricia Elizaquível

  2. Instituto de Agroquímica y Tecnología de Alimentos, Consejo Superior de Investigaciones Científicas, Madrid, Spain

    Rosa Aznar & Patricia Elizaquível

  3. Departamento de Biotecnología, Instituto de Agroquímica y Tecnología de los Alimentos, Apartado de Correos 73, 46100, Burjassot, Valencia, Spain

    Rosa Aznar

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  1. Rosa Aznar
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Correspondence to Rosa Aznar.

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Aznar, R., Elizaquível, P. Reliability of Listeria monocytogenes Identification by Specific PCR Assessed by Phenotypic and Genotypic Techniques. Food Anal. Methods 1, 243–251 (2008). https://doi.org/10.1007/s12161-008-9014-7

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