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Genotyping of Brucella abortus isolated in Gyeongbuk province by PFGE  

Jo, Min-Hee (Gyeongbuk Veterinary Service Laboratory)
Kim, Seong-Guk (Gyeongbuk Veterinary Service Laboratory)
Kim, Young-Hoan (Gyeongbuk Veterinary Service Laboratory)
Kim, Soon-Tae (Gyeongbuk Veterinary Service Laboratory)
Eom, Hyun-Jung (Gyeongbuk Veterinary Service Laboratory)
Jang, Young-Sui (Gyeongbuk Veterinary Service Laboratory)
Ko, Young-Hwal (Gyeongbuk Veterinary Service Laboratory)
Publication Information
Korean Journal of Veterinary Service / v.32, no.3, 2009 , pp. 257-264 More about this Journal
Abstract
Subtyping of Brucella abortus isolates is epidemiologically important for monitoring of bovine brucellosis outbreaks. Pulsed-field gel electrophoresis (PFGE) is considered as a gold standard of molecular typing methods to study the DNA polymorphisms of bacteria. In this study, we analyzed using PFGE the DNA fragment profiles of B. abortus isolated in Gyeongbuk province from 1998 to 2006. The genomic DNA was digested with the restriction endonuclease Xba I, Xho I and Smi I followed gel electrophoresis. No distinguishable patterns of the genomic DNA digested with Xba I and Xho I were observed among the field isolates of B. abortus tested in this study. But Smi I restriction enzyme resulted in two PFGE patterns consisting of 13-15 bands that ranged in size from 33 to 668bp by standard marker. The cluster analysis by DNA fingerprinting software showed 93.75% similarity between two PFGE patterns. No different PFGE patterns were recognized among the isolates originated from various years, regions and cow breeds.
Keywords
Brucella abortus; Molecular typing; PFGE;
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  • Reference
1 Beran GW. 1994. Handbook of zoonoses. 2nd Ed. CRC Press, Boca Raton: 9-39
2 Brenner DT, Kreig NR, Staley JT. 2005. Bergey's mannual of systematic bacteriology, 2nd Ed, Vol 2. Springer Press, New York: 370-389
3 Chain PSG, Comerci DJ, Tolmasky ME, Larimer FW, Malfatti SA, Vergez LM, Aguero F, Land ML, Ugalde RA, Garcia E. 2005. Whole-genome analyses of specia-tion events in pathogenic brucellae. lnfect lmmun 73(12): 8353-8361   DOI   ScienceOn
4 Cloeckaert A, Grayon M, Verger JM, Letesson JJ, Goifroid F 2000. Conservation of seven genes involved in the biosynthesis of the lipopolysaccharide O-side chain in Brucella spp. Res MicrobioI 151(3): 209-216   DOI   ScienceOn
5 Foster G, MacMillan AP, Godfroid J, Howie F, Ross HM, Cloeckaert A, Reid RJ, Brew S, Patterson IAP. 2002. A review of brucella sp. infection of sea mammals with particular emphasis on isolates from Scotland. Vet Microbiol 90: 563-580   DOI   ScienceOn
6 Verstreate DR, Winter AJ. 1984. Comparison of sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles and antigenic related-ness among outer membrane proteins of 49 Brucella abortus strains. Infect Immun 46(1): 182-187   PUBMED
7 Vizcaino N, Cloeckaert A, Verger J, Grayon M, FernandezLago L. 2000. DNA polymorphism in the genus brucella. Microbes Infect 2(9): 1089-1100   DOI   ScienceOn
8 Alton GG. 1975. Laboratory techniques in brucellosis, 2nd Ed. WHO, Geneva: 11-63
9 Halling SM, Peterson-Burch BD, Bricker BJ, Zuemer RL, Qing Z, Li LL, Kapur V, Alt DP, Olsδn SC. 2005. Completion of the genome sequence of Brucella abortus and comparison to the highly similar genomes of Brucella melitensis and Brucella suis. J Bacteriol 187(8): 2715-2726   DOI   ScienceOn
10 Moreno E, Stackebrandt E, Dorsch M, Wolters J, Busch M, Mayer H. 1990. Brucella abortus 16S rRNA and lipid A reveal a phylogenetic relationship with members of the alpha-2 subdivision of the class proteobacteria. J Bacteriol 172(7): 3569-3576   PUBMED
11 Olson MV. 1989. Separation of large DNA molecules by pulsed-field gel electrophoresis. A review of the basic phenomenology. J Chromator 470(2): 377-383   DOI   ScienceOn
12 Timoney JF, Gillespie JH, Scott FW, Barlough JE. 1988. Hagan and Bruner' s Microbiology and infectious disease of domestic animals, 8th Ed. Cornell University Press, New York. 135-152
13 Nielsen K, Duncan JR. 1990. Animal brucellosis, CRC Press, Boston. 1-453
14 Jensen AE, Cheville NF, Ewalt DR, Payeur JB, Thoen CO. 1995. Application of pulsed-field gel electrophoresis for differentiation of vaccine strain RB51 from field isolates of Brucella abortus from cattle, bison, and elk. Am J Vet Res 56(3): 308-312   PUBMED
15 CDC, Pulsenet. 2004. Standardized laboratory protocol for molecular subtyping of Escherchia coli O157:H7, non-typhoidal salmonella sero-types, and Shigella sonnei by PFGE. http://www.cdc.gov/pulsenet/protocols
16 DelVecchio VG, Kapatral V, Redkar RJ, Patra G, Mujer C, Los T, Ivanova N, Anderson I, Bha-ttacharyya A, Lykidis A, Reznik G, Jablonski L, Larsen N, D'Souza M, Bemal A, Mazur M, GoJtsman E, Selkov E, Elzer PH, Hagius S, 0 'Callaghan D, Letesson JJ, Haselkom R, Kyrpides N, Overbeek R. 2002. The genome sequence of the facultative intracellular patho-gen Brucella melitensis. PNAS 99(1): 443-448   DOI   ScienceOn
17 Hunter SB, Vauterin P, Lambert-Fair MA, Van Duyne MS, Kubota K, Graves L, Wrigley D, Barrett T, Ribot E. 2005. Establishment of a universal size standard strain for use with the pulsenet standardized pulsed-field gel electro-phoresis protocol: converting the national databases to the new size standard. J Clin Microbiol 43(3): 1045-1050   DOI   ScienceOn
18 Tenover FC, Arbeit RD, Goering RV, Mickelsen PA, Murray BE, Persing DH, Swaminathan B. 1995. Interpreting chromosomal DNA restric-tion patterns produced by pulsed-fieId gel electrophoresis: criteria for bacterial strain typing. J Clin MicrobioI 33(9): 2233-2239
19 Verger JM, Grimmont F, Grimmont PA, Grayon M. 1985. Taxonomy of genus brucella. Ann Ins Pasteur Microbiol 138(2): 235-238
20 Jensen AE, Cheville NF, Thoen CO, MacMillan AP, Miller WG. 1999. Genomic fingerprinting and development of a dendrogram for Brucella spp. isloated from seals, porpoises, and dolphins. J Vet Diagn lnvest 11: 152-157   DOI   ScienceOn
21 RidIer AL, Leyland MJ, Fenwick SG, West DM. 2005. Demonstration of polymorphism among Brucella ovis field isolates by pulsed-field gel electrophoresis. Vet microbiol l08: 69-74   DOI   ScienceOn
22 Moreno E, Cloeckaert A, Moriyon I. 2002. Brucella evolution and taxonomy. Vet Microbiol 90(1): 209-227   DOI   ScienceOn
23 Cloeckaert A, Verger JM, Grayon M, Grepinet O. 1995 Restriction site polymorphism of the genes encoding the major 25kDa and 36kDa outer-membrane proteins of brucella. Microbiology 141(9): 2111-2121   DOI   ScienceOn
24 Paulsen IT, Seshadri R, Nelson KE, Eisen JA, Heidelberg JF, Read TD, Dodson RJ, Umayam L, Brinkac LM, Beanan MJ, Daugherty SC, Deboy RT, Durkin AS, Kolonay JF, Madupu R, NeIson WC, Ayodeji B, Kraul M, Shetty J, Malek J, Van Aken SE, Riedmuller S, Tettelin H, Gill SR, White O, Salzberg SL, Hoover DL, LindIer LE, Halling SM, Boyle SM, Fraser CM. 2002. The Brucella suis genome reveals funda-mental similarities between animal and plant pathogens and symbionts. PNAS 99(20): 13148-13153   DOI   ScienceOn
25 Allardet-servent A, Bourg G, Ramuz M, Pages M, Bellis M, Roizes G. 1988. DNA polymorphism in strains of the genus brucella. J Bacterol 170(10): 4603-4607
26 Chu G, Vollrath D, Davis RW. 1986. Separation of large DNA molecules by contour-clamped homogeneous electric fieIds. Science 234(4783): 1582-1585   DOI   PUBMED
27 Ficht TA, Bearden SW, Sowa BA, Marquis H. 1990. Genetic variation at the omp2 porin locus of the brucellae: Species-specific mar-kers. Mol Microbiol 4(7): 1135-1142   DOI   ScienceOn
28 McPeek FD, Coyle-Morris JF, Gemmill RM. 1986. Separation of large DNA molecules by modified pulsed field gradient gel electro-phoresis. Anal Biochem 156(2): 274-285   DOI   ScienceOn
29 Jesen AE, Ewalt DR, Cheville NF, Thoen CO, Payeur JB. 1996. Determination of stability of Brucella abortus RB51 by use of genomic fingerprint， oxidative metabolism, and colonial morphology and differentiation of strain RB51 from B. abortus isolates from bison and elk. J Clin Microbiol 34(3): 628-633   PUBMED