Diversity of swine Bordetella bronchiseptica isolates evaluated by RAPD analysis and PFGE.
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.
Shin
Eun Kyung
EK
South Branch, Gangwon Veterinary Service, Wonju 200-170, Korea.
Seo
Yeon Soo
YS
Han
Jeong Hee
JH
Hahn
Tae Wook
TW
D000818:Animals; D016950:Bordetella bronchiseptica / Q000235:genetics*; D016000:Cluster Analysis; D017931:DNA Primers; D016521:Electrophoresis, Gel, Pulsed-Field; D014644:Genetic Variation; D007723:Korea; D019105:Random Amplified Polymorphic DNA Technique; D013045:Species Specificity; D013552:Swine / Q000382:microbiology*
D017931:DNA Primers