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

• Shin EK
• Seo YS
• Han JH
• Hahn TW

• KMID: 1104238
• DOI: http://doi.org/10.4142/jvs.2007.8.1.65

Abstract

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.

Keyword

Bordetella bronchiseptica; genetic diversity; PFGE; RAPD

MeSH Terms

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

Figure

• 

  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.

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