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J Vet Sci. 2007 Mar;8(1):65-73 |
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Diversity of swine Bordetella bronchiseptica isolates evaluated by RAPD analysis and PFGE
Eun-Kyung Shin1, Yeon-Soo Seo2, Jeong Hee Han2, Tae-Wook Hahn2,* |
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1South Branch, Gangwon Veterinary Service, Wonju 200-170, Korea
2School of Veterinary Medicine, Kangwon National University, Chuncheon 200-701, Korea
* twhahn@kangwon.ac.kr |
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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 ≥ 4 bands) and 11 A subtypes (differing by ≤ 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.
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