Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1996 Jan;34(1):15–19. doi: 10.1128/jcm.34.1.15-19.1996

Comparison of ribotyping, arbitrarily primed PCR, and pulsed-field gel electrophoresis for molecular typing of Listeria monocytogenes.

M Louie 1, P Jayaratne 1, I Luchsinger 1, J Devenish 1, J Yao 1, W Schlech 1, A Simor 1
PMCID: PMC228720  PMID: 8748263

Abstract

Fifty-one clinical isolates of Listeria monocytogenes (15 isolates from two outbreaks and 36 epidemiologically unrelated isolates) were typed by conventional serotyping, ribotyping (RT), pulsed-field gel electrophoresis (PFGE), and arbitrarily primed PCR (AP-PCR). Serotyping was unable to distinguish between related and unrelated strains of L. monocytogenes. Each of the three molecular methods showed excellent typeability and reproducibility. Restriction with EcoRI and PvuII gave 16 and 23 RT patterns, respectively. Restriction with ApaI or SmaI generated 22 and 26 PFGE profiles, respectively. ApaI profiles were easier to interpret, with 10 to 15 bands each, while SmaI profiles had 15 to 20 bands each. AP-PCR with two different primers yielded 29 and 31 randomly amplified polymorphic DNA patterns, respectively. Strains from the same outbreak shared concordant patterns by each of the three methods. Of the three techniques evaluated, RT was the least discriminating and could not distinguish between strains from the two outbreaks. The abilities of AP-PCR and PFGE to differentiate between strains were comparable. However, AP-PCR was more rapid and easier to perform. We conclude that the DNA profiles generated by either AP-PCR or PFGE can be used to differentiate outbreak strains from epidemiologically unrelated strains and to clearly identify unrelated strains as being distinct from one another. We recommend that at least two independent primers be used for AP-PCR typing in order to improve its discriminatory power.

Full Text

The Full Text of this article is available as a PDF (357.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Audurier A., Martin C. Phage typing of Listeria monocytogenes. Int J Food Microbiol. 1989 Jun;8(3):251–257. doi: 10.1016/0168-1605(89)90022-6. [DOI] [PubMed] [Google Scholar]
  2. Baloga A. O., Harlander S. K. Comparison of methods for discrimination between strains of Listeria monocytogenes from epidemiological surveys. Appl Environ Microbiol. 1991 Aug;57(8):2324–2331. doi: 10.1128/aem.57.8.2324-2331.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bannerman T. L., Hancock G. A., Tenover F. C., Miller J. M. Pulsed-field gel electrophoresis as a replacement for bacteriophage typing of Staphylococcus aureus. J Clin Microbiol. 1995 Mar;33(3):551–555. doi: 10.1128/jcm.33.3.551-555.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bibb W. F., Schwartz B., Gellin B. G., Plikaytis B. D., Weaver R. E. Analysis of Listeria monocytogenes by multilocus enzyme electrophoresis and application of the method to epidemiologic investigations. Int J Food Microbiol. 1989 Jun;8(3):233–239. doi: 10.1016/0168-1605(89)90018-4. [DOI] [PubMed] [Google Scholar]
  5. Boerlin P., Bannerman E., Ischer F., Rocourt J., Bille J. Typing Listeria monocytogenes: a comparison of random amplification of polymorphic DNA with 5 other methods. Res Microbiol. 1995 Jan;146(1):35–49. doi: 10.1016/0923-2508(96)80269-5. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Brosius J., Ullrich A., Raker M. A., Gray A., Dull T. J., Gutell R. R., Noller H. F. Construction and fine mapping of recombinant plasmids containing the rrnB ribosomal RNA operon of E. coli. Plasmid. 1981 Jul;6(1):112–118. doi: 10.1016/0147-619x(81)90058-5. [DOI] [PubMed] [Google Scholar]
  8. Farber J. M., Addison C. J. RAPD typing for distinguishing species and strains in the genus Listeria. J Appl Bacteriol. 1994 Sep;77(3):242–250. doi: 10.1111/j.1365-2672.1994.tb03070.x. [DOI] [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. Gellin B. G., Broome C. V. Listeriosis. JAMA. 1989 Mar 3;261(9):1313–1320. [PubMed] [Google Scholar]
  11. 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]
  12. Hunter P. R., Gaston M. A. Numerical index of the discriminatory ability of typing systems: an application of Simpson's index of diversity. J Clin Microbiol. 1988 Nov;26(11):2465–2466. doi: 10.1128/jcm.26.11.2465-2466.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. MacGowan A. P., O'Donaghue K., Nicholls S., McLauchlin J., Bennett P. M., Reeves D. S. Typing of Listeria spp. by random amplified polymorphic DNA (RAPD) analysis. J Med Microbiol. 1993 May;38(5):322–327. doi: 10.1099/00222615-38-5-322. [DOI] [PubMed] [Google Scholar]
  15. Mazurier S. I., Audurier A., Marquet-Van der Mee N., Notermans S., Wernars K. A comparative study of randomly amplified polymorphic DNA analysis and conventional phage typing for epidemiological studies of Listeria monocytogenes isolates. Res Microbiol. 1992 Jun;143(5):507–512. doi: 10.1016/0923-2508(92)90097-8. [DOI] [PubMed] [Google Scholar]
  16. Mazurier S. I., Wernars K. Typing of Listeria strains by random amplification of polymorphic DNA. Res Microbiol. 1992 Jun;143(5):499–505. doi: 10.1016/0923-2508(92)90096-7. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Nocera D., Altwegg M., Martinetti Lucchini G., Bannerman E., Ischer F., Rocourt J., Bille J. Characterization of Listeria strains from a foodborne listeriosis outbreak by rDNA gene restriction patterns compared to four other typing methods. Eur J Clin Microbiol Infect Dis. 1993 Mar;12(3):162–169. doi: 10.1007/BF01967106. [DOI] [PubMed] [Google Scholar]
  19. Nørrung B., Gerner-Smidt P. Comparison of multilocus enzyme electrophoresis (MEE), ribotyping, restriction enzyme analysis (REA) and phage typing for typing of Listeria monocytogenes. Epidemiol Infect. 1993 Aug;111(1):71–79. doi: 10.1017/s0950268800056697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. Welsh J., McClelland M. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res. 1990 Dec 25;18(24):7213–7218. doi: 10.1093/nar/18.24.7213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Williams J. G., Kubelik A. R., Livak K. J., Rafalski J. A., Tingey S. V. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 1990 Nov 25;18(22):6531–6535. doi: 10.1093/nar/18.22.6531. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES