This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Bricker, B J Articles by Halling, S M Search for Related Content PubMed PubMed Citation Articles by Bricker, B J Articles by Halling, S M

Differentiation of Brucella abortus bv. 1, 2, and 4, Brucella melitensis, Brucella ovis, and Brucella suis bv. 1 by PCR.

National Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Ames, Iowa 50010.

ABSTRACT

Several PCR assays which identify the genus Brucella but do not discriminate among species have been reported. We describe a PCR assay that comprises five oligonucleotide primers which can identify selected biovars of four species of Brucella. Individual biovars within a species are not differentiated. The assay can identify three biovars (1, 2, and 4) of B. abortus, all three biovars of B. melitensis, biovar 1 of B. suis, and all B. ovis biovars. These biovars include all of the Brucella species typically isolated from cattle in the United States, a goal of the present research. The assay exploits the polymorphism arising from species-specific localization of the genetic element IS711 in the Brucella chromosome. Identity is determined by the size(s) of the product(s) amplified from primers hybridizing at various distances from the element. The performance of the assay with U.S. field isolates was highly effective. When 107 field isolates were screened by the described method, there was 100% agreement with the identifications made by conventional methods. Six closely related bacteria (Agrobacterium radiobacter, Agrobacterium rhizogenes, Ochrobactrum anthropi, Rhizobium leguminosarum, Rhizobium meliloti, and Rhodospirillum rubrum) and two control bacteria (Bordetella bronchiseptica and Escherichia coli) tested negative by the assay.

This article has been cited by other articles:

• Scholz, H. C., Hubalek, Z., Sedlacek, I., Vergnaud, G., Tomaso, H., Al Dahouk, S., Melzer, F., Kampfer, P., Neubauer, H., Cloeckaert, A., Maquart, M., Zygmunt, M. S., Whatmore, A. M., Falsen, E., Bahn, P., Gollner, C., Pfeffer, M., Huber, B., Busse, H.-J., Nockler, K. (2008). Brucella microti sp. nov., isolated from the common vole Microtus arvalis. Int. J. Syst. Evol. Microbiol. 58: 375-382 [Abstract] [Full Text]  
• De, B. K., Stauffer, L., Koylass, M. S., Sharp, S. E., Gee, J. E., Helsel, L. O., Steigerwalt, A. G., Vega, R., Clark, T. A., Daneshvar, M. I., Wilkins, P. P., Whatmore, A. M. (2008). Novel Brucella Strain (BO1) Associated with a Prosthetic Breast Implant Infection. J. Clin. Microbiol. 46: 43-49 [Abstract] [Full Text]  
• Foster, J. T., Okinaka, R. T., Svensson, R., Shaw, K., De, B. K., Robison, R. A., Probert, W. S., Kenefic, L. J., Brown, W. D., Keim, P. (2008). Real-Time PCR Assays of Single-Nucleotide Polymorphisms Defining the Major Brucella Clades. J. Clin. Microbiol. 46: 296-301 [Abstract] [Full Text]  
• Garcia-Yoldi, D., Le Fleche, P., De Miguel, M. J., Munoz, P. M., Blasco, J. M., Cvetnic, Z., Marin, C. M., Vergnaud, G., Lopez-Goni, I. (2007). Comparison of Multiple-Locus Variable-Number Tandem-Repeat Analysis with Other PCR-Based Methods for Typing Brucella suis Isolates. J. Clin. Microbiol. 45: 4070-4072 [Abstract] [Full Text]  
• Scott, J. C., Koylass, M. S., Stubberfield, M. R., Whatmore, A. M. (2007). Multiplex Assay Based on Single-Nucleotide Polymorphisms for Rapid Identification of Brucella Isolates at the Species Level. Appl. Environ. Microbiol. 73: 7331-7337 [Abstract] [Full Text]  
• Stoffregen, W. C., Olsen, S. C., Jack Wheeler, C., Bricker, B. J., Palmer, M. V., Jensen, A. E., Halling, S. M., Alt, D. P. (2007). Diagnostic Characterization of a Feral Swine Herd Enzootically Infected with Brucella. jvdi 19: 227-237 [Abstract] [Full Text]  
• Whatmore, A. M., Shankster, S. J., Perrett, L. L., Murphy, T. J., Brew, S. D., Thirlwall, R. E., Cutler, S. J., MacMillan, A. P. (2006). Identification and Characterization of Variable-Number Tandem-Repeat Markers for Typing of Brucella spp.. J. Clin. Microbiol. 44: 1982-1993 [Abstract] [Full Text]  
• Wellinghausen, N., Nockler, K., Sigge, A., Bartel, M., Essig, A., Poppert, S. (2006). Rapid Detection of Brucella spp. in Blood Cultures by Fluorescence In Situ Hybridization.. J. Clin. Microbiol. 44: 1828-1830 [Abstract] [Full Text]  
• Ulrich, M. P., Norwood, D. A., Christensen, D. R., Ulrich, R. L. (2006). Using real-time PCR to specifically detect Burkholderia mallei.. J Med Microbiol 55: 551-559 [Abstract] [Full Text]  
• Christensen, D. R., Hartman, L. J., Loveless, B. M., Frye, M. S., Shipley, M. A., Bridge, D. L., Richards, M. J., Kaplan, R. S., Garrison, J., Baldwin, C. D., Kulesh, D. A., Norwood, D. A. (2006). Detection of Biological Threat Agents by Real-Time PCR: Comparison of Assay Performance on the R.A.P.I.D., the LightCycler, and the Smart Cycler Platforms. Clin. Chem. 52: 141-145 [Abstract] [Full Text]  
• Al-Nakkas, A., Mustafa, A. S., Wright, S. G (2005). Large-scale evaluation of a single-tube nested PCR for the laboratory diagnosis of human brucellosis in Kuwait. J Med Microbiol 54: 727-730 [Abstract] [Full Text]  
• Adone, R., Ciuchini, F., Marianelli, C., Tarantino, M., Pistoia, C., Marcon, G., Petrucci, P., Francia, M., Riccardi, G., Pasquali, P. (2005). Protective Properties of Rifampin-Resistant Rough Mutants of Brucella melitensis. Infect. Immun. 73: 4198-4204 [Abstract] [Full Text]  
• Halling, S. M., Peterson-Burch, B. D., Bricker, B. J., Zuerner, R. L., Qing, Z., Li, L.-L., Kapur, V., Alt, D. P., Olsen, S. C. (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: 2715-2726 [Abstract] [Full Text]  
• Whatmore, A. M., Murphy, T. J., Shankster, S., Young, E., Cutler, S. J., Macmillan, A. P. (2005). Use of Amplified Fragment Length Polymorphism To Identify and Type Brucella Isolates of Medical and Veterinary Interest. J. Clin. Microbiol. 43: 761-769 [Abstract] [Full Text]  
• Newby, D. T., Hadfield, T. L., Roberto, F. F. (2003). Real-Time PCR Detection of Brucella abortus: a Comparative Study of SYBR Green I, 5'-Exonuclease, and Hybridization Probe Assays. Appl. Environ. Microbiol. 69: 4753-4759 [Abstract] [Full Text]  
• Motiwala, A. S., Strother, M., Amonsin, A., Byrum, B., Naser, S. A., Stabel, J. R., Shulaw, W. P., Bannantine, J. P., Kapur, V., Sreevatsan, S. (2003). Molecular Epidemiology of Mycobacterium avium subsp. paratuberculosis: Evidence for Limited Strain Diversity, Strain Sharing, and Identification of Unique Targets for Diagnosis. J. Clin. Microbiol. 41: 2015-2026 [Abstract] [Full Text]  
• Bardenstein, S., Mandelboim, M., Ficht, T. A., Baum, M., Banai, M. (2002). Identification of the Brucella melitensis Vaccine Strain Rev.1 in Animals and Humans in Israel by PCR Analysis of the PstI Site Polymorphism of Its omp2 Gene. J. Clin. Microbiol. 40: 1475-1480 [Abstract] [Full Text]  
• Gándara, B., Merino, A. L., Rogel, M. A., Martínez-Romero, E. (2001). Limited Genetic Diversity of Brucella spp.. J. Clin. Microbiol. 39: 235-240 [Abstract] [Full Text]  
• Cloeckaert, A., Grayon, M., Grepinet, O. (2000). An IS711 Element Downstream of the bp26 Gene Is a Specific Marker of Brucella spp. Isolated from Marine Mammals. CVI 7: 835-839 [Abstract] [Full Text]  
• Ewalt, D. R., Bricker, B. J. (2000). Validation of the Abbreviated Brucella AMOS PCR as a Rapid Screening Method for Differentiation of Brucella abortus Field Strain Isolates and the Vaccine Strains, 19 and RB51. J. Clin. Microbiol. 38: 3085-3086 [Abstract] [Full Text]  
• Sreevatsan, S., Bookout, J. B., Ringpis, F., Perumaalla, V. S., Ficht, T. A., Adams, L. G., Hagius, S. D., Elzer, P. H., Bricker, B. J., Kumar, G. K., Rajasekhar, M., Isloor, S., Barathur, R. R. (2000). A Multiplex Approach to Molecular Detection of Brucella abortus and/or Mycobacterium bovis Infection in Cattle. J. Clin. Microbiol. 38: 2602-2610 [Abstract] [Full Text]  
• Bricker, B. J., Ewalt, D. R., MacMillan, A. P., Foster, G., Brew, S. (2000). Molecular Characterization of Brucella Strains Isolated from Marine Mammals. J. Clin. Microbiol. 38: 1258-1262 [Abstract] [Full Text]  
• Vemulapalli, R., McQuiston, J. R., Schurig, G. G., Sriranganathan, N., Halling, S. M., Boyle, S. M. (1999). Identification of an IS711 Element Interrupting the wboA Gene of Brucella abortus Vaccine Strain RB51 and a PCR Assay To Distinguish Strain RB51 from Other Brucella Species and Strains. CVI 6: 760-764 [Abstract] [Full Text]  
• Vizcaino, N., Cloeckaert, A., Zygmunt, M. S., Fernandez-Lago, L. (1999). Molecular Characterization of a Brucella Species Large DNA Fragment Deleted in Brucella abortus Strains: Evidence for a Locus Involved in the Synthesis of a Polysaccharide. Infect. Immun. 67: 2700-2712 [Abstract] [Full Text]  
• Fox, K. F., Fox, A., Nagpal, M., Steinberg, P., Heroux, K. (1998). Identification of Brucella by Ribosomal-Spacer-Region PCR and Differentiation of Brucella canis from Other Brucella spp. Pathogenic for Humans by Carbohydrate Profiles. J. Clin. Microbiol. 36: 3217-3222 [Abstract] [Full Text]