PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of aemPermissionsJournals.ASM.orgJournalAEM ArticleJournal InfoAuthorsReviewers
 
Appl Environ Microbiol. 1997 August; 63(8): 2961–2965.
PMCID: PMC168593

Evaluation of luciferase reporter bacteriophage A511::luxAB for detection of Listeria monocytogenes in contaminated foods.

Abstract

A511::luxAB is a recombinant derivative of a broad-host-range bacteriophage specific for the genus Listeria, transducing bacterial bioluminescence into infected cells. In this study, we have evaluated its use for rapid and easy testing of contaminated foods and environmental samples for the presence of viable Listeria cells, in comparison to the standard plating procedure. With a short preenrichment step of 20 h, the system was capable of detecting very low initial contamination rates in several foods artificially contaminated with Listeria monocytogenes Scott A cells. In ricotta cheese, chocolate pudding, and cabbage, less than one cell per g of food could be clearly identified by comparing the light emission of phage-infected samples to that of controls without lux phage. In foods having a large and complex microbial background flora, such as minced meat and soft cheese, at least 10 cells per g were necessary to produce a positive bioluminescence signal. Of 348 potentially contaminated natural food and environmental samples, 55 were found to be Listeria positive by the lux phage method. The standard plating procedure detected 57 positive samples. Some differences were observed with respect to the individual samples, i.e., the lux phage procedure detected more positive samples among the dairy products and environmental samples, whereas the plating procedure revealed more contaminated meat and poultry samples. Overall, both methods performed similarly, i.e., were equally sensitive. However, the minimum time required for detection of Listeria with the luciferase phage assay was 24 h, which is much shorter than the 4 days needed by the standard plating method. Furthermore, a most probable number technique with three parallels, based on the use of A511::luxAB for differentiation of positive and negative tubes, is described. The method enables rapid enumeration of low levels of Listeria cells in several foods tested, against the background of a competing microflora.

Full Text

The Full Text of this article is available as a PDF (792K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bessesen MT, Luo QA, Rotbart HA, Blaser MJ, Ellison RT., 3rd Detection of Listeria monocytogenes by using the polymerase chain reaction. Appl Environ Microbiol. 1990 Sep;56(9):2930–2932. [PMC free article] [PubMed]
  • Bickley J, Short JK, McDowell DG, Parkes HC. Polymerase chain reaction (PCR) detection of Listeria monocytogenes in diluted milk and reversal of PCR inhibition caused by calcium ions. Lett Appl Microbiol. 1996 Feb;22(2):153–158. [PubMed]
  • Bubert A, Köhler S, Goebel W. The homologous and heterologous regions within the iap gene allow genus- and species-specific identification of Listeria spp. by polymerase chain reaction. Appl Environ Microbiol. 1992 Aug;58(8):2625–2632. [PMC free article] [PubMed]
  • Bubert A, Schubert P, Köhler S, Frank R, Goebel W. Synthetic peptides derived from the Listeria monocytogenes p60 protein as antigens for the generation of polyclonal antibodies specific for secreted cell-free L. monocytogenes p60 proteins. Appl Environ Microbiol. 1994 Sep;60(9):3120–3127. [PMC free article] [PubMed]
  • Curtis GD, Lee WH. Culture media and methods for the isolation of Listeria monocytogenes. Int J Food Microbiol. 1995 Jun;26(1):1–13. [PubMed]
  • Emond E, Fliss I, Pandian S. A ribosomal DNA fragment of Listeria monocytogenes and its use as a genus-specific probe in an aqueous-phase hybridization assay. Appl Environ Microbiol. 1993 Aug;59(8):2690–2697. [PMC free article] [PubMed]
  • Farber JM, Peterkin PI. Listeria monocytogenes, a food-borne pathogen. Microbiol Rev. 1991 Sep;55(3):476–511. [PMC free article] [PubMed]
  • Fliss I, St Laurent M, Emond E, Simard RE, Lemieux R, Ettriki A, Pandian S. Anti-DNA.RNA antibodies: an efficient tool for non-isotopic detection of Listeria species through a liquid-phase hybridization assay. Appl Microbiol Biotechnol. 1995 Aug-Sep;43(4):717–724. [PubMed]
  • Hof H, Rocourt J. Is any strain of Listeria monocytogenes detected in food a health risk? Int J Food Microbiol. 1992 Jul;16(3):173–182. [PubMed]
  • Loessner MJ. Improved procedure for bacteriophage typing of Listeria strains and evaluation of new phages. Appl Environ Microbiol. 1991 Mar;57(3):882–884. [PMC free article] [PubMed]
  • Loessner MJ, Bell RH, Jay JM, Shelef LA. Comparison of seven plating media for enumeration of Listeria spp. Appl Environ Microbiol. 1988 Dec;54(12):3003–3007. [PMC free article] [PubMed]
  • Loessner MJ, Busse M. Bacteriophage typing of Listeria species. Appl Environ Microbiol. 1990 Jun;56(6):1912–1918. [PMC free article] [PubMed]
  • Loessner MJ, Rees CE, Stewart GS, Scherer S. Construction of luciferase reporter bacteriophage A511::luxAB for rapid and sensitive detection of viable Listeria cells. Appl Environ Microbiol. 1996 Apr;62(4):1133–1140. [PMC free article] [PubMed]
  • Loessner MJ, Scherer S. Organization and transcriptional analysis of the Listeria phage A511 late gene region comprising the major capsid and tail sheath protein genes cps and tsh. J Bacteriol. 1995 Nov;177(22):6601–6609. [PMC free article] [PubMed]
  • Starbuck MA, Hill PJ, Stewart GS. Ultra sensitive detection of Listeria monocytogenes in milk by the polymerase chain reaction (PCR). Lett Appl Microbiol. 1992 Dec;15(6):248–252. [PubMed]
  • Uyttendaele M, Schukkink R, van Gemen B, Debevere J. Development of NASBA, a nucleic acid amplification system, for identification of Listeria monocytogenes and comparison to ELISA and a modified FDA method. Int J Food Microbiol. 1995 Sep;27(1):77–89. [PubMed]
  • Wendlinger G, Loessner MJ, Scherer S. Bacteriophage receptors on Listeria monocytogenes cells are the N-acetylglucosamine and rhamnose substituents of teichoic acids or the peptidoglycan itself. Microbiology. 1996 Apr;142(Pt 4):985–992. [PubMed]
  • Wernars K, Heuvelman CJ, Chakraborty T, Notermans SH. Use of the polymerase chain reaction for direct detection of Listeria monocytogenes in soft cheese. J Appl Bacteriol. 1991 Feb;70(2):121–126. [PubMed]
  • Zink R, Loessner MJ. Classification of virulent and temperate bacteriophages of Listeria spp. on the basis of morphology and protein analysis. Appl Environ Microbiol. 1992 Jan;58(1):296–302. [PMC free article] [PubMed]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)