Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1996 Mar;34(3):641–646. doi: 10.1128/jcm.34.3.641-646.1996

Evaluation and comparison of molecular techniques for epidemiological typing of Salmonella enterica subsp. enterica serovar dublin.

B Liebisch 1, S Schwarz 1
PMCID: PMC228862  PMID: 8904430

Abstract

A total of 28 unrelated isolates of the Salmonella enterica subsp. enterica serovar dublin (S. dublin) collected during a 6-year period, as well as four samples of the S. dublin live vaccine strain Bovisaloral and its prototype strain S. dublin 442/039, were investigated by different molecular typing methods for the following reasons: (i) to find the most discriminatory method for the epidemiological typing of isolates belonging to this Salmonella serovar and (ii) to evaluate these methods for their capacity to discriminate among the live vaccine strain Bovisaloral, its prototype strain S. dublin 442/039, and field isolates of the serovar dublin. Five different plasmid profiles were observed; a virulence plasmid of 76 kbp as identified by hybridization with an spvB-spvC gene probe was present in all isolates. The detection of 16S rRNA genes and that of IS200 elements proved to be unsuitable for the epidemiological typing of S. dublin; only one hybridization pattern could be observed with each of these methods. The results obtained from macrorestriction analysis strongly depended on the choice of restriction enzyme. While the enzyme NotI yielded the lowest discriminatory index among all enzymes tested, it was the only enzyme that allowed discrimination between the Bovisaloral vaccine strain and its prototype strain. In contrast to the enzymes XbaI and SpeI, which only differentiated among the S. dublin field isolates, XhoI as well as AvrII also produced restriction fragment patterns of the Bovisaloral strain and of its prototype strain that were not shared by any of the S. dublin field isolates. Macrorestriction analysis proved to be the most discriminatory method not only for the epidemiological typing of S. dublin field isolates but also for the identification of the S. dublin live vaccine strain Bovisaloral.

Full Text

The Full Text of this article is available as a PDF (372.1 KB).

Selected References

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

  1. Beyer W., Geue L. Characterization of the virulence regions in the plasmids of three live Salmonella vaccines. Zentralbl Bakteriol. 1992 Jun;277(1):10–21. doi: 10.1016/s0934-8840(11)80865-0. [DOI] [PubMed] [Google Scholar]
  2. Browning L. M., Wray C., Platt D. J. Diversity and molecular variation among plasmids in Salmonella enterica serotype Dublin based on restriction enzyme fragmentation pattern analysis. Epidemiol Infect. 1995 Apr;114(2):237–248. doi: 10.1017/s0950268800057903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chowdry N., Threlfall E. J., Rowe B., Stanley J. Genotype analysis of faecal and blood isolates of Salmonella dublin from humans in England and Wales. Epidemiol Infect. 1993 Apr;110(2):217–225. doi: 10.1017/s0950268800068138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Esteban E., Snipes K., Hird D., Kasten R., Kinde H. Use of ribotyping for characterization of Salmonella serotypes. J Clin Microbiol. 1993 Feb;31(2):233–237. doi: 10.1128/jcm.31.2.233-237.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fang F. C., Fierer J. Human infection with Salmonella dublin. Medicine (Baltimore) 1991 May;70(3):198–207. doi: 10.1097/00005792-199105000-00004. [DOI] [PubMed] [Google Scholar]
  6. Gibert I., Barbé J., Casadesús J. Distribution of insertion sequence IS200 in Salmonella and Shigella. J Gen Microbiol. 1990 Dec;136(12):2555–2560. doi: 10.1099/00221287-136-12-2555. [DOI] [PubMed] [Google Scholar]
  7. Gulig P. A. Virulence plasmids of Salmonella typhimurium and other salmonellae. Microb Pathog. 1990 Jan;8(1):3–11. doi: 10.1016/0882-4010(90)90003-9. [DOI] [PubMed] [Google Scholar]
  8. Hoiseth S. K., Stocker B. A. Aromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines. Nature. 1981 May 21;291(5812):238–239. doi: 10.1038/291238a0. [DOI] [PubMed] [Google Scholar]
  9. Hormaeche C. E., Joysey H. S., Desilva L., Izhar M., Stocker B. A. Immunity conferred by Aro- Salmonella live vaccines. Microb Pathog. 1991 Feb;10(2):149–158. doi: 10.1016/0882-4010(91)90075-l. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Iglesias A., Ceglowski P., Trautner T. A. Plasmid transformation in Bacillus subtilis. Effects of the insertion of Bacillus subtilis rRNA genes into plasmids. Mol Gen Genet. 1983;192(1-2):149–154. doi: 10.1007/BF00327660. [DOI] [PubMed] [Google Scholar]
  12. Kado C. I., Liu S. T. Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol. 1981 Mar;145(3):1365–1373. doi: 10.1128/jb.145.3.1365-1373.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lax A. J., Pullinger G. D., Baird G. D., Williamson C. M. The virulence plasmid of Salmonella dublin: detailed restriction map and analysis by transposon mutagenesis. J Gen Microbiol. 1990 Jun;136(6):1117–1123. doi: 10.1099/00221287-136-6-1117. [DOI] [PubMed] [Google Scholar]
  14. Lester A., Eriksen N. H., Nielsen H., Nielsen P. B., Friis-Møller A., Bruun B., Scheibel J., Gaarslev K., Kolmos H. J. Non-typhoid Salmonella bacteraemia in Greater Copenhagen 1984 to 1988. Eur J Clin Microbiol Infect Dis. 1991 Jun;10(6):486–490. doi: 10.1007/BF01963934. [DOI] [PubMed] [Google Scholar]
  15. Linde K. Herstellung von stabilen Salmonella-Impfstämmen durch Kopplung von zwei unabhängig voneinander nichtvermehrungsbegrenzenden attenuierenden Markern. Arch Exp Veterinarmed. 1980;34(1):19–32. [PubMed] [Google Scholar]
  16. Macrina F. L., Kopecko D. J., Jones K. R., Ayers D. J., McCowen S. M. A multiple plasmid-containing Escherichia coli strain: convenient source of size reference plasmid molecules. Plasmid. 1978 Jun;1(3):417–420. doi: 10.1016/0147-619x(78)90056-2. [DOI] [PubMed] [Google Scholar]
  17. Millemann Y., Lesage M. C., Chaslus-Dancla E., Lafont J. P. Value of plasmid profiling, ribotyping, and detection of IS200 for tracing avian isolates of Salmonella typhimurium and S. enteritidis. J Clin Microbiol. 1995 Jan;33(1):173–179. doi: 10.1128/jcm.33.1.173-179.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Montenegro M. A., Morelli G., Helmuth R. Heteroduplex analysis of Salmonella virulence plasmids and their prevalence in isolates of defined sources. Microb Pathog. 1991 Dec;11(6):391–397. doi: 10.1016/0882-4010(91)90035-9. [DOI] [PubMed] [Google Scholar]
  19. Olsen J. E. An improved method for rapid isolation of plasmid DNA from wild-type gram-negative bacteria for plasmid restriction profile analysis. Lett Appl Microbiol. 1990 May;10(5):209–212. doi: 10.1111/j.1472-765x.1990.tb01335.x. [DOI] [PubMed] [Google Scholar]
  20. Olsen J. E., Brown D. J., Baggesen D. L., Bisgaard M. Biochemical and molecular characterization of Salmonella enterica serovar berta, and comparison of methods for typing. Epidemiol Infect. 1992 Apr;108(2):243–260. doi: 10.1017/s0950268800049724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Olsen J. E., Brown D. J., Skov M. N., Christensen J. P. Bacterial typing methods suitable for epidemiological analysis. Applications in investigations of salmonellosis among livestock. Vet Q. 1993 Dec;15(4):125–135. doi: 10.1080/01652176.1993.9694390. [DOI] [PubMed] [Google Scholar]
  22. Powell N. G., Threlfall E. J., Chart H., Rowe B. Subdivision of Salmonella enteritidis PT 4 by pulsed-field gel electrophoresis: potential for epidemiological surveillance. FEMS Microbiol Lett. 1994 Jun 1;119(1-2):193–198. doi: 10.1111/j.1574-6968.1994.tb06888.x. [DOI] [PubMed] [Google Scholar]
  23. Schwarz S., Liebisch B. Pulsed-field gel electrophoretic identification of Salmonella enterica serovar Typhimurium live vaccine strain Zoosaloral H. Lett Appl Microbiol. 1994 Dec;19(6):469–472. doi: 10.1111/j.1472-765x.1994.tb00984.x. [DOI] [PubMed] [Google Scholar]
  24. Schwarz S., Liebisch B. Use of ribotyping, IS200 typing and plasmid analysis for the identification of Salmonella enterica subsp. enterica serovar Typhimurium vaccine strain Zoosaloral H and its differentiation from wild type strains of the same serovar. Zentralbl Bakteriol. 1994 Nov;281(4):442–450. doi: 10.1016/s0934-8840(11)80330-0. [DOI] [PubMed] [Google Scholar]
  25. Segall T., Lindberg A. A. Salmonella dublin experimental infection in calves: protection after oral immunization with an auxotrophic aroA live vaccine. Zentralbl Veterinarmed B. 1991 Mar;38(2):142–160. doi: 10.1111/j.1439-0450.1991.tb00857.x. [DOI] [PubMed] [Google Scholar]
  26. Sigwart D. F., Stocker B. A., Clements J. D. Effect of a purA mutation on efficacy of Salmonella live-vaccine vectors. Infect Immun. 1989 Jun;57(6):1858–1861. doi: 10.1128/iai.57.6.1858-1861.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stanley J., Baquar N., Threlfall E. J. Genotypes and phylogenetic relationships of Salmonella typhimurium are defined by molecular fingerprinting of IS200 and 16S rrn loci. J Gen Microbiol. 1993 Jun;139(Pt 6):1133–1140. doi: 10.1099/00221287-139-6-1133. [DOI] [PubMed] [Google Scholar]
  28. Stanley J., Burnens A. P., Threlfall E. J., Chowdry N., Goldsworthy M. Genetic relationships among strains of Salmonella enteritidis in a national epidemic in Switzerland. Epidemiol Infect. 1992 Apr;108(2):213–220. doi: 10.1017/s0950268800049694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stanley J., Chowdry N., Powell N., Threlfall E. J. Chromosomal genotypes (evolutionary lines) of Salmonella berta. FEMS Microbiol Lett. 1992 Aug 15;74(2-3):247–252. doi: 10.1016/0378-1097(92)90437-s. [DOI] [PubMed] [Google Scholar]
  30. Taylor D. N., Bied J. M., Munro J. S., Feldman R. A. Salmonella dublin infections in the United States, 1979-1980. J Infect Dis. 1982 Sep;146(3):322–327. doi: 10.1093/infdis/146.3.322. [DOI] [PubMed] [Google Scholar]
  31. Thong K. L., Cheong Y. M., Puthucheary S., Koh C. L., Pang T. Epidemiologic analysis of sporadic Salmonella typhi isolates and those from outbreaks by pulsed-field gel electrophoresis. J Clin Microbiol. 1994 May;32(5):1135–1141. doi: 10.1128/jcm.32.5.1135-1141.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Thong K. L., Ngeow Y. F., Altwegg M., Navaratnam P., Pang T. Molecular analysis of Salmonella enteritidis by pulsed-field gel electrophoresis and ribotyping. J Clin Microbiol. 1995 May;33(5):1070–1074. doi: 10.1128/jcm.33.5.1070-1074.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Threlfall E. J., Frost J. A. The identification, typing and fingerprinting of Salmonella: laboratory aspects and epidemiological applications. J Appl Bacteriol. 1990 Jan;68(1):5–16. doi: 10.1111/j.1365-2672.1990.tb02542.x. [DOI] [PubMed] [Google Scholar]
  34. Threlfall E. J., Hall M. L., Rowe B. Salmonella bacteraemia in England and Wales, 1981-1990. J Clin Pathol. 1992 Jan;45(1):34–36. doi: 10.1136/jcp.45.1.34. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES