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. 1997 Jul;65(7):2707–2716. doi: 10.1128/iai.65.7.2707-2716.1997

Ribotypes and virulence gene polymorphisms suggest three distinct Listeria monocytogenes lineages with differences in pathogenic potential.

M Wiedmann 1, J L Bruce 1, C Keating 1, A E Johnson 1, P L McDonough 1, C A Batt 1
PMCID: PMC175382  PMID: 9199440

Abstract

A total of 133 Listeria monocytogenes isolates were characterized by ribotyping and allelic analysis of the virulence genes hly, actA, and inlA to uncover linkages between independent phylogenetic and specific virulence markers. PCR-restriction fragment length polymorphisms revealed 8 hly, 11 inl4, and 2 actA alleles. The combination of these virulence gene alleles and ribotype patterns separated L. monocytogenes into three distinct lineages. While distinct hly and inlA alleles were generally found to cluster into these three lineages, actA alleles segregated independently. These three phylogenetic lineages were confirmed when 22 partial actA DNA sequences were analyzed. The clinical history of the L. monocytogenes strains showed evidence for differences in pathogenic potential among the three lineages. Lineage I contains all strains isolated during epidemic outbreaks of listeriosis, while no human isolates were found in lineage III. Animal isolates were found in all three lineages. We found evidence that isolates from lineages I and III have a higher plaquing efficiency than lineage II strains in a cell culture assay. Strains from lineage III also seem to form larger plaques than strains from lineage II. A distinctive ribotype fragment and unique 16S rRNA gene sequences furthermore suggest that lineage III might represent a L. monocytogenes subspecies. None of the 20 human isolates available but 11% of our animal isolates were grouped in this lineage, indicating that strains in this lineage might have reduced virulence for humans.

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Selected References

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  1. Brosch R., Chen J., Luchansky J. B. Pulsed-field fingerprinting of listeriae: identification of genomic divisions for Listeria monocytogenes and their correlation with serovar. Appl Environ Microbiol. 1994 Jul;60(7):2584–2592. doi: 10.1128/aem.60.7.2584-2592.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bruce J. L., Hubner R. J., Cole E. M., McDowell C. I., Webster J. A. Sets of EcoRI fragments containing ribosomal RNA sequences are conserved among different strains of Listeria monocytogenes. Proc Natl Acad Sci U S A. 1995 May 23;92(11):5229–5233. doi: 10.1073/pnas.92.11.5229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brundage R. A., Smith G. A., Camilli A., Theriot J. A., Portnoy D. A. Expression and phosphorylation of the Listeria monocytogenes ActA protein in mammalian cells. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11890–11894. doi: 10.1073/pnas.90.24.11890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chakraborty T., Ebel F., Wehland J., Dufrenne J., Notermans S. Naturally occurring virulence-attenuated isolates of Listeria monocytogenes capable of inducing long term protection against infection by virulent strains of homologous and heterologous serotypes. FEMS Immunol Med Microbiol. 1994 Nov;10(1):1–9. doi: 10.1111/j.1574-695X.1994.tb00004.x. [DOI] [PubMed] [Google Scholar]
  5. Collins M. D., Wallbanks S., Lane D. J., Shah J., Nietupski R., Smida J., Dorsch M., Stackebrandt E. Phylogenetic analysis of the genus Listeria based on reverse transcriptase sequencing of 16S rRNA. Int J Syst Bacteriol. 1991 Apr;41(2):240–246. doi: 10.1099/00207713-41-2-240. [DOI] [PubMed] [Google Scholar]
  6. Czajka J., Batt C. A. Verification of causal relationships between Listeria monocytogenes isolates implicated in food-borne outbreaks of listeriosis by randomly amplified polymorphic DNA patterns. J Clin Microbiol. 1994 May;32(5):1280–1287. doi: 10.1128/jcm.32.5.1280-1287.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Czajka J., Bsat N., Piani M., Russ W., Sultana K., Wiedmann M., Whitaker R., Batt C. A. Differentiation of Listeria monocytogenes and Listeria innocua by 16S rRNA genes and intraspecies discrimination of Listeria monocytogenes strains by random amplified polymorphic DNA polymorphisms. Appl Environ Microbiol. 1993 Jan;59(1):304–308. doi: 10.1128/aem.59.1.304-308.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Domann E., Wehland J., Rohde M., Pistor S., Hartl M., Goebel W., Leimeister-Wächter M., Wuenscher M., Chakraborty T. A novel bacterial virulence gene in Listeria monocytogenes required for host cell microfilament interaction with homology to the proline-rich region of vinculin. EMBO J. 1992 May;11(5):1981–1990. doi: 10.1002/j.1460-2075.1992.tb05252.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Farber J. M., Peterkin P. I. Listeria monocytogenes, a food-borne pathogen. Microbiol Rev. 1991 Sep;55(3):476–511. doi: 10.1128/mr.55.3.476-511.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fleming D. W., Cochi S. L., MacDonald K. L., Brondum J., Hayes P. S., Plikaytis B. D., Holmes M. B., Audurier A., Broome C. V., Reingold A. L. Pasteurized milk as a vehicle of infection in an outbreak of listeriosis. N Engl J Med. 1985 Feb 14;312(7):404–407. doi: 10.1056/NEJM198502143120704. [DOI] [PubMed] [Google Scholar]
  11. Friederich E., Gouin E., Hellio R., Kocks C., Cossart P., Louvard D. Targeting of Listeria monocytogenes ActA protein to the plasma membrane as a tool to dissect both actin-based cell morphogenesis and ActA function. EMBO J. 1995 Jun 15;14(12):2731–2744. doi: 10.1002/j.1460-2075.1995.tb07274.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gerstel B., Gröbe L., Pistor S., Chakraborty T., Wehland J. The ActA polypeptides of Listeria ivanovii and Listeria monocytogenes harbor related binding sites for host microfilament proteins. Infect Immun. 1996 Jun;64(6):1929–1936. doi: 10.1128/iai.64.6.1929-1936.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Graves L. M., Swaminathan B., Reeves M. W., Hunter S. B., Weaver R. E., Plikaytis B. D., Schuchat A. Comparison of ribotyping and multilocus enzyme electrophoresis for subtyping of Listeria monocytogenes isolates. J Clin Microbiol. 1994 Dec;32(12):2936–2943. doi: 10.1128/jcm.32.12.2936-2943.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gutekunst K. A., Holloway B. P., Carlone G. M. DNA sequence heterogeneity in the gene encoding a 60-kilodalton extracellular protein of Listeria monocytogenes and other Listeria species. Can J Microbiol. 1992 Aug;38(8):865–870. doi: 10.1139/m92-141. [DOI] [PubMed] [Google Scholar]
  15. Ho J. L., Shands K. N., Friedland G., Eckind P., Fraser D. W. An outbreak of type 4b Listeria monocytogenes infection involving patients from eight Boston hospitals. Arch Intern Med. 1986 Mar;146(3):520–524. [PubMed] [Google Scholar]
  16. Hubner R. J., Cole E. M., Bruce J. L., McDowell C. I., Webster J. A. Types of Listeria monocytogenes predicted by the positions of EcoRI cleavage sites relative to ribosomal RNA sequences. Proc Natl Acad Sci U S A. 1995 May 23;92(11):5234–5238. doi: 10.1073/pnas.92.11.5234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jacquet C., Catimel B., Brosch R., Buchrieser C., Dehaumont P., Goulet V., Lepoutre A., Veit P., Rocourt J. Investigations related to the epidemic strain involved in the French listeriosis outbreak in 1992. Appl Environ Microbiol. 1995 Jun;61(6):2242–2246. doi: 10.1128/aem.61.6.2242-2246.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lennon D., Lewis B., Mantell C., Becroft D., Dove B., Farmer K., Tonkin S., Yeates N., Stamp R., Mickleson K. Epidemic perinatal listeriosis. Pediatr Infect Dis. 1984 Jan-Feb;3(1):30–34. doi: 10.1097/00006454-198401000-00008. [DOI] [PubMed] [Google Scholar]
  19. Linnan M. J., Mascola L., Lou X. D., Goulet V., May S., Salminen C., Hird D. W., Yonekura M. L., Hayes P., Weaver R. Epidemic listeriosis associated with Mexican-style cheese. N Engl J Med. 1988 Sep 29;319(13):823–828. doi: 10.1056/NEJM198809293191303. [DOI] [PubMed] [Google Scholar]
  20. McLauchlin J. Distribution of serovars of Listeria monocytogenes isolated from different categories of patients with listeriosis. Eur J Clin Microbiol Infect Dis. 1990 Mar;9(3):210–213. doi: 10.1007/BF01963840. [DOI] [PubMed] [Google Scholar]
  21. Nørrung B., Skovgaard N. Application of multilocus enzyme electrophoresis in studies of the epidemiology of Listeria monocytogenes in Denmark. Appl Environ Microbiol. 1993 Sep;59(9):2817–2822. doi: 10.1128/aem.59.9.2817-2822.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Piffaretti J. C., Kressebuch H., Aeschbacher M., Bille J., Bannerman E., Musser J. M., Selander R. K., Rocourt J. Genetic characterization of clones of the bacterium Listeria monocytogenes causing epidemic disease. Proc Natl Acad Sci U S A. 1989 May;86(10):3818–3822. doi: 10.1073/pnas.86.10.3818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Quentin R., Huet H., Wang F. S., Geslin P., Goudeau A., Selander R. K. Characterization of Streptococcus agalactiae strains by multilocus enzyme genotype and serotype: identification of multiple virulent clone families that cause invasive neonatal disease. J Clin Microbiol. 1995 Oct;33(10):2576–2581. doi: 10.1128/jcm.33.10.2576-2581.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rasmussen O. F., Beck T., Olsen J. E., Dons L., Rossen L. Listeria monocytogenes isolates can be classified into two major types according to the sequence of the listeriolysin gene. Infect Immun. 1991 Nov;59(11):3945–3951. doi: 10.1128/iai.59.11.3945-3951.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rasmussen O. F., Skouboe P., Dons L., Rossen L., Olsen J. E. Listeria monocytogenes exists in at least three evolutionary lines: evidence from flagellin, invasive associated protein and listeriolysin O genes. Microbiology. 1995 Sep;141(Pt 9):2053–2061. doi: 10.1099/13500872-141-9-2053. [DOI] [PubMed] [Google Scholar]
  26. Rodrigues J., Scaletsky I. C., Campos L. C., Gomes T. A., Whittam T. S., Trabulsi L. R. Clonal structure and virulence factors in strains of Escherichia coli of the classic serogroup O55. Infect Immun. 1996 Jul;64(7):2680–2686. doi: 10.1128/iai.64.7.2680-2686.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schlech W. F., 3rd, Lavigne P. M., Bortolussi R. A., Allen A. C., Haldane E. V., Wort A. J., Hightower A. W., Johnson S. E., King S. H., Nicholls E. S. Epidemic listeriosis--evidence for transmission by food. N Engl J Med. 1983 Jan 27;308(4):203–206. doi: 10.1056/NEJM198301273080407. [DOI] [PubMed] [Google Scholar]
  28. Schuchat A., Swaminathan B., Broome C. V. Epidemiology of human listeriosis. Clin Microbiol Rev. 1991 Apr;4(2):169–183. doi: 10.1128/cmr.4.2.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schwartz B., Hexter D., Broome C. V., Hightower A. W., Hirschhorn R. B., Porter J. D., Hayes P. S., Bibb W. F., Lorber B., Faris D. G. Investigation of an outbreak of listeriosis: new hypotheses for the etiology of epidemic Listeria monocytogenes infections. J Infect Dis. 1989 Apr;159(4):680–685. doi: 10.1093/infdis/159.4.680. [DOI] [PubMed] [Google Scholar]
  30. Smith G. A., Marquis H., Jones S., Johnston N. C., Portnoy D. A., Goldfine H. The two distinct phospholipases C of Listeria monocytogenes have overlapping roles in escape from a vacuole and cell-to-cell spread. Infect Immun. 1995 Nov;63(11):4231–4237. doi: 10.1128/iai.63.11.4231-4237.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Vines A., Reeves M. W., Hunter S., Swaminathan B. Restriction fragment length polymorphism in four virulence-associated genes of Listeria monocytogenes. Res Microbiol. 1992 Mar-Apr;143(3):281–294. doi: 10.1016/0923-2508(92)90020-o. [DOI] [PubMed] [Google Scholar]
  32. Yang Y., Gabriel D. W. Intragenic recombination of a single plant pathogen gene provides a mechanism for the evolution of new host specificities. J Bacteriol. 1995 Sep;177(17):4963–4968. doi: 10.1128/jb.177.17.4963-4968.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Zhang Q., Wise K. S. Molecular basis of size and antigenic variation of a Mycoplasma hominis adhesin encoded by divergent vaa genes. Infect Immun. 1996 Jul;64(7):2737–2744. doi: 10.1128/iai.64.7.2737-2744.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. d'Hauteville H., Sansonetti P. J. Phosphorylation of IcsA by cAMP-dependent protein kinase and its effect on intracellular spread of Shigella flexneri. Mol Microbiol. 1992 Apr;6(7):833–841. doi: 10.1111/j.1365-2958.1992.tb01534.x. [DOI] [PubMed] [Google Scholar]

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