tailieunhanh - Báo cáo khoa học: " Diversity of swine Bordetella bronchiseptica isolates evaluated by RAPD analysis and PFGE"

Tuyển tập các báo cáo nghiên cứu khoa học quốc tế về bệnh thú y đề tài: Diversity of swine Bordetella bronchiseptica isolates evaluated by RAPD analysis and PFGE | J. Vet. Sci. 2007 8 1 65-73 JOURNAL OF Veterinary Science Diversity of swine Bordetella bronchiseptica isolates evaluated by RAPD analysis and PFGE Eun-Kyung Shin1 Yeon-Soo Seo2 Jeong Hee Han2 Tae-Wook Hahn2 1 South Branch Gangwon Veterinary Service Wonju 200-170 Korea 2School of Veterinary Medicine Kangwon National University Chuncheon 200-701 Korea 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 the pattern produced by OPA-08 was shared by 26 isolates and the pattern produced by OPA-18 was shared by 40 isolates . 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 Xbal produced a total of 15 patterns consisting of 4 PFGE types A B Bl and c differing by 4 bands and 11A 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. Key words Bordetella bronchiseptica genetic diversity PFGE RAPD Introduction Bordetella are Gram-negative bacteria that cause respiratory tract infections in humans and animals. Species in the genus Bordetella are close phenotypically possess common antigens and share a high degree of DNA similarity 3 . .