J Vet Sci.  2007 Mar;8(1):65-73. 10.4142/jvs.2007.8.1.65.

Diversity of swine Bordetella bronchiseptica isolates evaluated by RAPD analysis and PFGE

  • 1South Branch, Gangwon Veterinary Service, Wonju 200-170, Korea.
  • 2School of Veterinary Medicine, Kangwon National University, Chuncheon 200-701, Korea. twhahn@kangwon.ac.kr


The degree of genetic diversity in 45 Bordetella (B.) bronchiseptica strains comprised of a vaccine strain (N = 1), reference strains (N = 3) and field isolates (N = 41) was evaluated using random amplified polymorphic DNA (RAPD) fingerprinting and pulsed-field gel electrophoresis (PFGE). Three candidate primers were selected for RAPD analysis after screening 20 random decamer oligonucleotides for their discriminatory abilities. The OPA-07, OPA-08 and OPA-18 primers yielded 10, 10, and 6 distinct fingerprint patterns, respectively. The most common identical RAPD pattern was produced by OPA-07 which was shared by 32 isolates (71.1%), the pattern produced by OPA-08 was shared by 26 isolates (57.8%), and the pattern produced by OPA-18 was shared by 40 isolates (88.9%). The RAPD patterns of the vaccine strain and the 3 reference strains did not match any of the patterns produced by the field isolates when primers OPA-07 and OPA-08 were used. PFGE using the restriction endonuclease XbaI produced a total of 15 patterns consisting of 4 PFGE types (A, B, B1 and C, differing by > or = 4 bands) and 11 A subtypes (differing by < or = 3 bands). Most of the field isolates exhibited identical type A and B patterns, suggesting that they were related. The vaccine strain and the three reference strains showed different PFGE patterns as compared to the identical type A strains.


Bordetella bronchiseptica; genetic diversity; PFGE; RAPD

MeSH Terms

Bordetella bronchiseptica/*genetics
Cluster Analysis
DNA Primers
Electrophoresis, Gel, Pulsed-Field
*Genetic Variation
Random Amplified Polymorphic DNA Technique
Species Specificity


  • Fig. 1 RAPD patterns and associated dendrograms of B. bronchiseptica strains. (A) RAPD patterns and associated dendrograms of B. bronchiseptica strains generated using primer OPA-07. (B) RAPD patterns and associated dendrograms of B. bronchiseptica strains generated using primer OPA-08. (C) RAPD patterns and associated dendrograms of B. bronchiseptica strains generated using primer OPA-18.

  • Fig. 2 Schematic magnification of PFGE patterns and dendrogram of XbaI PFGE patterns of B. bronchiseptica performed according to UPGMA. (A) Lanes: M, lambda DNA standard marker; A~C, B. bronchiseptica PFGE patterns (B) PFGE patterns and associated dendrograms of B. bronchiseptica strains generated by digestion with XbaI.


1. Barbier N, Saulnier P, Chachaty E, Dumontier S, Andremont A. Random amplified polymorphic DNA typing versus pulsed-field gel electrophoresis for epidemiological typing of vancomycin-resistant enterococci. J Clin Microbiol. 1996. 34:1096–1099.
2. Baysinger A. PRDC: is it new or déjá vu? Pork '99. 1999. 19:64.
3. Bemis DA, Burns EH. Gyles CL, Thoen CO, editors. Bordetella. Pathogenesis of Bacterial Infections in Animals. 1993. 2nd ed. Ames: Iowa State University Press;201–215.
4. Bemis DA, Greisen HA, Appel MJ. Bacteriological variation among Bordetella bronchiseptica isolates from dogs and other species. J Clin Microbiol. 1977. 5:471–480.
5. Bemis DA, Plotkin BJ. Hemagglutination by Bordetella bronchiseptica. J Clin Microbiol. 1982. 15:1120–1127.
6. Binns SH, Speakman AJ, Dawson S, Bennett M, Gaskell RM, Hart CA. The use of pulsed-field gel electrophoresis to examine the epidemiology of Bordetella bronchiseptica isolated from cats and other species. Epidemiol Infect. 1998. 120:201–208.
7. Chang N, Chui L. A standardized protocol for the rapid preparation of bacterial DNA for pulsed-field gel electrophoresis. Diagn Microbiol Infect Dis. 1998. 31:275–279.
8. Cowan ST, Steel KJ. Manual for the Identification of Medical Bacterial. 1974. 2nd ed. London: Cambridge University Press;89–90.
9. Gautom RK. Rapid pulsed-field gel electrophoresis protocol for typing of Escherichia coli O157:H7 and other gramnegative organisms in 1 day. J Clin Microbiol. 1997. 35:2977–2980.
10. Giles CJ. Leman AD, Straw BE, Mengeling WL, D'Allaire S, Taylor DJ, editors. Bordetellosis. Disease of Swine. 1992. 7th ed. Ames: Iowa State University Press;436–445.
11. Graham AC, Abruzzo GK. Occurrence and characterization of plasmids in field isolates of Bordetella bronchiseptica. Am J Vet Res. 1982. 43:1852–1855.
12. Grimont F, Grimont PA. Ribosomal ribonucleic acid gene restriction patterns as potential taxonomic tools. Ann Inst Pasteur Microbiol. 1986. 137B:165–175.
13. Harris DL, Switzer WP. Turbinate atrophy in young pigs exposed to Bordetella bronchiseptica, Pasteurella multocida, and combined inoculum. Am J Vet Res. 1968. 29:777–785.
14. Johnson R, Sneath PHA. Taxonomy of Bordetella and related organisms of the families Achromobacteraceae, Brucellaeae and Neisseriaceae. Int J Syst Bacteriol. 1973. 23:381–404.
15. Keil DJ, Fenwick B. Evaluation of canine Bordetella bronchiseptica isolates using randomly amplified polymorphic DNA fingerprinting and ribotyping. Vet Microbiol. 1999. 66:41–51.
16. Khattak MN, Matthews RC. Genetic relatedness of Bordetella species as determined by macrorestriction digests resolved by pulsed-field gel electrophoresis. Int J Syst Bacteriol. 1993. 43:659–664.
17. Moissenet D, Valcin M, Marchand V, Grimprel E, Begue P, Garbarg-Chenon A, Vu-Thien H. Comparative DNA analysis of Bordetella pertussis clinical isolates by pulsed-field gel electrophoresis, randomly amplified polymorphism DNA, and ERIC polymerase chain reaction. FEMS Microbiol Lett. 1996. 143:127–132.
18. Musser JM, Hewlett EL, Peppler MS, Selander RK. Genetic diversity and relationships in populations of Bordetella spp. J Bacteriol. 1986. 166:230–237.
19. Pedersen KB. The serology of Bordetella bronchiseptica isolated from pigs compared with strains from other animal species. Acta Pathol Microbiol Scand Suppl. 1975. 83:590–594.
20. Peppler MS, Schrumpf ME. Phenotypic variation and modulation in Bordetella bronchiseptica. Infect Immun. 1984. 44:681–687.
21. Register KB, Boisvert A, Ackermann MR. Use of ribotyping to distinguish Bordetella bronchiseptica isolates. Int J Syst Bacteriol. 1997. 47:678–683.
22. Ross RF, Switzer WP, Duncan JR. Comparison of pathogenicity of various isolates of Bordetella bronchiseptica in young pigs. Can J Comp Med Vet Sci. 1967. 31:53–57.
23. Rutter JM, Collings LA. Pedersen KB, Nielsen NC, editors. The virulence of Bordetella bronchiseptica in atrophic rhinitis of pigs. Atrophic Rhinitis in Pigs. 1983. Luxembourg: Commission of the European Communities;77–83.
24. Sacco RE, Register KB, Nordholm GE. Restriction endonuclease analysis discriminates Bordetella bronchiseptica isolates. J Clin Microbiol. 2000. 38:4387–4393.
25. Seo YS, Lee SH, Shin EK, Kim SJ, Jung R, Hahn TW. Pulsed-field gel electrophoresis genotyping of Salmonella gallinarum and comparison with random amplified polymorphic DNA. Vet Microbiol. 2006. 115:349–357.
26. Smith IM, Baskerville AJ. A selective medium facilitating the isolation and recognition the Bordetella bronchiseptica in pigs. Res Vet Sci. 1979. 27:187–192.
27. Stull TL, LiPuma JJ, Edlind TD. A broad-spectrum probe for molecular epidemiology of bacteria: ribosomal RNA. J Infect Dis. 1988. 157:280–286.
28. Tenover FC, Arbeit RD, Goering RV, Mickelsen PA, Murray BE, Persing DH, Swaminathan B. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: Criteria for bacterial strain typing. J Clin Microbiol. 1995. 33:2233–2239.
29. Tynkkynen S, Satokari R, Saarela M, Mattila-Sandholm T, Saxelin M. Comparison of ribotyping, randomly amplified polymorphic DNA analysis, and pulsed-field gel electrophoresis in typing of Lactobacillus rhamnosus and L. casei strains. Appl Environ Microbiol. 1999. 65:3908–3914.
30. Vu-Thien H, Dulot C, Moissenet D, Fauroux B, Garbarg-Chenon A. Comparison of randomly amplified polymorphic DNA analysis and pulsed-field gel electrophoresis for typing of Moraxella catarrhalis strains. J Clin Microbiol. 1999. 37:450–452.
31. Winstanley C, Shina A, Dawson S, Gaskell RM, Hart CA. Variation in Bordetella bronchiseptica flaA does not correlate with typing by macro-restriction analysis by pulsed-field gel electrophoresis. J Med Microbiol. 2001. 50:255–260.
32. Yuk MH, Heininger U, Martinez de Tejada G, Miller JF. Human but not ovine isolates of Bordetella parapertussis are highly clonal as determined by PCR-based RAPD fingerprinting. Infection. 1998. 26:270–273.
33. Zhang Y, Rajagopalan M, Brown BA, Wallace RJ Jr. Randomly amplified polymorphic DNA PCR for comparison of Mycobacterium abscessus strains from nosocomial outbreaks. J Clin Microbiol. 1997. 35:3132–3139.
Full Text Links
  • JVS
export Copy
  • Twitter
  • Facebook
Similar articles
Copyright © 2023 by Korean Association of Medical Journal Editors. All rights reserved.     E-mail: koreamed@kamje.or.kr