Current Methods of Human and Animal Brucellosis Diagnostics

DOI: 10.4236/aid.2013.33026   PDF   HTML   XML   6,024 Downloads   10,827 Views   Citations


Brucellosis is an urgent infectious disease of livestock and wild animals and the commonest human zoonosis. Diagnosis of brucellosis is rather complicated and it has to be obligatorily confirmed by laboratory testing. Direct bacteriological and molecular methods and indirect serological tests are used for brucellosis diagnostics. The choice of the diagnostic tools depends on the overall epidemiological situation in the region and the objectives of the study: validation of the diagnosis, screening (monitoring), cross-sectional studies or confirmation of brucellosis-free status of the region. The review describes current bacteriological, serological and molecular methods, routinely used for the diagnosis of brucellosis in humans and animals. The perspectives of brucellosis diagnostics are also discussed.

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E. Smirnova, A. Vasin, N. Sandybaev, S. Klotchenko, M. Plotnikova, O. Chervyakova, A. Sansyzbay and O. Kiselev, "Current Methods of Human and Animal Brucellosis Diagnostics," Advances in Infectious Diseases, Vol. 3 No. 3, 2013, pp. 177-184. doi: 10.4236/aid.2013.33026.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] J. Godfroid, K. Nielsen and C. Saegerman, “Diagnosis of Brucellosis in Livestock and Wildlife,” Croatian Medical Journal, Vol. 51, No. 4, 2010, pp. 296-305. doi:10.3325/cmj.2010.51.296
[2] P. P. Poester, K. Nielsen, L. E. Samartino and W. L. Yu, “Diagnosis of Brucellosis,” The Open Veterinary Science Journal, Vol. 4, 2010, pp. 46-60.
[3] M. J. Corbel, “Brucellosis: An Overview,” Proceedings of the 1st International Conference on Emerging Zoonoses: Emerging Infectious Diseases, Jerusalem, AprilJune 1997, pp. 213-221.
[4] World Health Organization, “Fact Sheet N173,” World Health Organization, Geneva, 1997.
[5] G. Pappas, P. Papadimitriou, N. Akritidis, L. Christou and E. V. Tsianos, “The New Global Map of Human Brucellosis,” Lancet Infectious Diseases, Vol. 6, No. 2, 2006, pp. 91-99. doi:10.1016/S1473-3099(06)70382-6
[6] Office International des épizooties, “Manual of Standards for Diagnostic Tests and Vaccines,” Office International des épizooties, Paris, 2000.
[7] CDC, “Public Health Consequences of a False-Positive Laboratory Test Result for Brucella—Florida, Georgia, and Michigan, 2005,” Morbidity and Mortality Weekly Report (MMWR), Vol. 57, No. 22, 2008, pp. 603-605.
[8] P. Yagupsky, “Detection of Brucellae in Blood Cultures,” Journal of Clinical Microbiology, Vol. 37, No. 11, 1999, pp. 3437-3442.
[9] S. R. Porter, G. Czaplicki, J. Mainil, R. Guattéo, C. Saegerman and Q. Fever, “Current State of Knowledge and Perspectives of Research of a Neglected Zoonosis,” International Journal of Microbiology, Vol. 2011, 2011, Article ID: 248418. doi:10.1155/2011/248418
[10] G. Pappas, N. Akritidis, M. Bosilkovski and E. Tsianos, “Brucellosis,” The New England Journal of Medicine, Vol. 352, No. 22, 2005, pp. 2325-2336. doi:10.1056/NEJMra050570
[11] J.-M. Verger, F. Grimont, P. A. D. Grimont and M. Grayon, “Brucella, a Monospecific Genus as Shown by Deoxyribonucleic Acid Hybridization,” International Journal of Systematic Bacteriology, Vol. 35, No. 3, 1985, pp. 292-295. doi:10.1099/00207713-35-3-292
[12] W. L. Yu and K. Nielsen, “Review of Detection of Brucella spp. by Polymerase Chain Reaction,” Croatian Medical Journal, Vol. 51, No. 4, 2010, pp. 306-313. doi:10.3325/cmj.2010.51.306
[13] M. M. Baddour and D. H. Alkhalifa, “Evaluation of Three Polymerase CHAIN Reaction Techniques for Detection of Brucella DNA in Peripheral Human Blood,” Canadian Journal of Microbiology, Vol. 54, No. 5, 2008, pp. 352-357. doi:10.1139/W08-017
[14] L. A. Dauphin, R. J. Hutchins, L. A. Bost and M. D. Bowen, “Evaluation of Automated and Manual Commercial DNA Extraction Methods for Recovery of Brucella DNA from Suspensions and Spiked Swabs,” Journal of Clinical Microbiology, Vol. 47, No. 12, 2009, pp. 3920-3926. doi:10.1128/JCM.01288-09
[15] S. Ouahrani-Bettach, M. P. Soubrier and J. P. Liautard, “IS6501-Anchored PCR for the Detection and Identification of Brucella Species and Strains,” Journal of Applied Bacteriology, Vol. 81, No. 2, 1996, pp. 154-160. doi:10.1111/j.1365-2672.1996.tb04493.x
[16] K. Imaoka, M. Kimura, M. Suzuki, T. Kamiyama and A. Yamada, “Simultaneous Detection of the Genus Brucella by Combinatorial PCR,” Japanese Journal of Infectious Diseases, Vol. 60, No. 2-3, 2007, pp. 137-139.
[17] V. Hinic, I. Broadard, A. Thomann, Z. Cvetnic, P. V. Mkaya, J. Frey and C. Abril, “Novel Identification and Differentiation of Brucella melitensis, B. abortus, B. suis, B. ovis, B. canis, and B. neotmae Suitable for Both Conventional and Real-Time PCR Systems,” Journal of Microbiological Methods, Vol. 75, No. 2, 2008, pp. 375-378. doi:10.1016/j.mimet.2008.07.002
[18] D. S. Leal-Klevezas, I. O. Martinez-Vazquez, J. Garcia-Cantu, A. Lopez-Merina and J. P. Martinez-Soriano, “Use of Polymerase Chain Reaction to Detect Brucellaabortus Biovar 1 in Infected Goats,” Veterinary Microbiology, Vol. 75, No. 1, 2000, pp. 91-97. doi:10.1016/S0378-1135(00)00200-5
[19] F. J. Sangari and J. Aguero, “Identification of Brucella abortus B19 Vaccine Strain by the Detection of DNA Polymorphism at the ery Locus,” Vaccine, Vol. 12, No. 5, 1994, pp. 435-438. doi:10.1016/0264-410X(94)90121-X
[20] R. Vemulapalli, J. R. McQuiston, G. G. Shurig, N. Srirangnathan, S. M. Halling and S. M. Boyle, “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,” Clinical and Diagnostic Laboratory Immunology, Vol. 6, No. 5, 1999, pp. 760-764.
[21] B. J. Bricker and S. M. Halling, “Differentiation of Brucella abortus bv. 1, 2 and 4, Brucella melitensis, Brucella ovis, and Brucella suis bv. 1 by PCR,” Journal of Clinical Microbiology, Vol. 32, No. 11, 1994, pp. 2660-2666.
[22] D. R. Ewalt, “Screening Method for Differentiation of Brucella abortus Field Strain Isolates and Vaccine Strains, 19 and RB51,” Journal of Clinical Microbiology, Vol. 38, No. 8, 2000, pp. 3085-3086.
[23] A. A. Ocampo-Sosa, J. Agüero-Balbin and J. M. Garcia-Lobo, “Development of a New PCR Assay to Identify Brucella abortus Biovars 5, 6 and 9 and New Subgroup 3b of Biovar 3,” Veterinary Microbiology, Vol. 110, No. 1-2, 2005, pp. 41-51.
[24] W. L. Yu and K. Nielsen, “Review of Detection of Brucella spp. by Polymerase Chain Reaction,” Croatian Medical Journal, Vol. 51, No. 4, 2010, pp. 306-313. doi:10.3325/cmj.2010.51.306
[25] B. B. Alarcón, B. Vicedo and R. Aznar, “PCR-Based Procedures for Detection of Staphylococcus aureus and Their Application in Food,” Journal of Applied Microbiology, Vol. 100, No. 2, 2006, pp. 352-364. doi:10.1111/j.1365-2672.2005.02768.x
[26] M. M. Kattar, P. A. Zallouab, G. F. Araja, J. Samaha-Kfourya, H. Shbakloe, S. S. Kanjb, S. Khalifea and M. Deebc, “Development and Evaluation of Real-Time Polymerase Chain Reaction Assays on Whole Blood and Paraffin-Embedded Tissues for Rapid Diagnosis of Human Brucellosis,” Diagnostic Microbiology and Infectious Disease, Vol. 59, No. 1, 2007, pp. 23-32. doi:10.1016/j.diagmicrobio.2007.04.002
[27] B. J. Bricker, D. R. Ewalt and S. M. Halling, “Brucella ‘HOOF-Prints’: Strain Typing by Multi-Locus Analysis of Variable Number Tandem Repeats (VNTRs),” BMC Microbiology, Vol. 3, 2003, p. 15. doi:10.1186/1471-2180-3-15
[28] F. Le Fleche, I. Jacques, M. Grayon, S. Al Dahouk, P. Bouchon, F. Denoeud, K. Nockler, H. Neubauer, L. A. Guilloteau and G. Vergnaud, “Evaluation and Selection of Tandem Repeat Loci for a Brucella MLVA Typing Assay,” BMC Microbiology, Vol. 6, 2006, p. 9. doi:10.1186/1471-2180-6-9
[29] G. G. Alton, L. M. Jones, R. D. Angus and J. M. Verger, “Techniques for the Brucellosis Laboratory,” Institute National de la Recherche Agronomique, Paris, 1988.
[30] K. H. Nielsen, “Diagnosis of Brucellosis by Serology,” Veterinary Microbiology, Vol. 90, No. 1-4, 2002, pp. 447-459.
[31] W. Hill, “Standardization of the Complement Fixation test for Brucellosis,” Bull OIE, Vol. 60, 1963, pp. 401-410.
[32] I. Dohoo, P. Wright, G. Ruckerbauer, B. Samagh, F. Robertson and L. Forbes, “A Comparison of Five Serological Tests for Bovine Brucellosis,” The Canadian Journal of Veterinary Research, Vol. 50, No. 4, 1986, pp. 485-493.
[33] X. Rojas and O. Alonso, “ELISA for the Diagnosis and Epidemiology of Brucella abortus Infection in Cattle in Chile,” IAEA TECDOC, Vienna, 1994, pp. 77-82.
[34] L. Samartino, R. Gregoret, D. Gall and K. Nielsen, “Fluorescence Polarization Assay: Application to the Diagnosis of Bovine Brucellosis in Argentina,” Journal of Immunoassay, Vol. 20, No. 3, 1999, pp. 115-120. doi:10.1080/01971529909349347
[35] P. Pardon, R. Sanchis, G. Molenat, J. Marly and D. Renard, “Serological and Allergic Reactions of Ewes after Simultaneous Vaccinations with Two Living Attenuated Strains of Brucella and Salmonella,” Annales de Recherches Veterinaires, Vol. 21, No. 2, 1990, pp. 153-160.
[36] P. M. Munoz, C. M. Marín, D. Monreal, D. González, B. Garin-Bastuji, R. Díaz, R. C. Mainar-Jaime, I. Moriyón and J. M. Blasco, “Efficacy of Several Serological Tests and Antigens for Diagnosis of Bovine Brucellosis in the Presence of False-Positive Serological Results Due to Yersinia enterocolitica O:9,” Clinical and Vaccine Immunology, Vol. 12, No. 1, 2005, pp. 141-151. doi:10.1128/CDLI.12.1.141-151.2005
[37] V. Weynants, A. Tibor, P. A. Denoel, C. Saegerman, J. Godfroid, P. Thiange and J. J. Letesson, “Infection of Cattle with Yersinia enterocolitica O:9 a Cause of the False Positive Serological Reactions in Bovine Brucellosis Diagnostic Tests,” Veterinary Microbiology, Vol. 48, No. 1-2, 1996, pp. 101-112. doi:10.1016/0378-1135(95)00153-0
[38] K. Nielsen, P. Smith, J. Widdison, D. Gall, L. Kelly, W. Kelly and P. Nicoletti, “Serological Relationship between Cattle Exposed to Brucella abortus, Yersinia enterocolitica O:9 and Escherichia coli O157:H7,” Veterinary Microbiology, Vol. 100, No. 1-2, 2004, pp. 25-30. doi:10.1016/j.vetmic.2003.12.010
[39] K. H. Nielsen, L. Kelly, D. Gall, P. Nicoletti and W. Kelly, “Improved Competitive Enzyme Immunoassay for the Diagnosis of Bovine Brucelliosis,” Veterinary Immunology and Immunopathology, Vol. 46, No. 3-4, 1995, pp. 285-291. doi:10.1016/0165-2427(94)05361-U
[40] V. Weynants, D. Gilson, A. Cloeckaert, P. A. Denoel, A. Tibor, P. Thiange, J. N. Limet and J. J. Letesson, “Characterization of a Monoclonal Antibody Specific for Brucella Smooth Lipopolysaccharide and Development of a Competitive Enzyme-Linked Immunosorbent Assay to Improve the Serological Diagnosis of Brucellosis,” Clinical and Diagnostic Laboratory Immunology, Vol. 3, No. 3, 1996, pp. 309-314.
[41] J. A. McGiven, J. D. Tucker, L. L. Perrett, J. A. Stack, S. D. Brew and A. P. MacMillan, “Validation of FPA and cELISA for the Detection of Antibodies to Brucella abortus in Cattle Sera and Comparison to SAT, CFT, and iELISA,” Journal of Immunological Methods, Vol. 278, No. 1-2, 2003, pp. 171-178. doi:10.1016/S0022-1759(03)00201-1
[42] J. E. Mayfield, B. J. Bricker, H. Godfrey, R. M. Crosby, D. J. Knight, S. M. Halling, D. Balinsky and L. B. Tabatabai, “The Cloning, Expression, and Nucleotide Sequence of a Gene Coding for an Immunogenic Brucella abortus Protein,” Gene, Vol. 63, No. 1, 1988, pp. 1-9. doi:10.1016/0378-1119(88)90540-9
[43] B. J. Bricker, L. B. Tabatabai, B. L. Deyoe and J. E. Mayfield, “Conservation of Antigenicity in a 31-kDa Brucella Protein,” Veterinary Microbiology, Vol. 18, No. 3-4, 1988, pp. 313-325. doi:10.1016/0378-1135(88)90096-X
[44] L. Zhang, X. A. Wu, F. L. Zhang, C. H. An, Y. X. Sun, W. T. Bai and Z. K. Xu, “Soluble Expression and Purification of Brucella Cell Surface Protein (BCSP31) of Brucella melitensis and Preparation of Anti-BCSP31 Monoclonal Antibodies,” Molecular Biology Reports, Vol. 39, No. 1, 2012, pp. 431-438. doi:10.1007/s11033-011-0755-9
[45] O. L. Rossetti, A. I. Arese, M. L. Boschiroli and S. L. Cravero, “Cloning of Brucella abortus Gene and Characterization of Expressed 26-Kilodalton Periplasmic Protein: Potential Use for Diagnosis,” Journal of Clinical Microbiology, Vol. 34, No. 1, 1996, pp. 165-169.
[46] A. Cloeckaert, H. Salih-AljDebbarh, N. Vizcaino, E. Saman, G. Dubray and M. S. Zygmunt, “Cloning, Nucleotide Sequence, and Expression of the Brucella melitensis bp26 Gene Coding for a Protein Immunogenic in Infected Sheep,” FEMS Microbiology Letters, Vol. 140, No. 2-3, 1996, pp. 139-144. doi:10.1111/j.1574-6968.1996.tb08327.x
[47] L. E. Lindler, T. L. Hadfield, B. D. Tall, N. J. Snellings, F. A. Rubin, L. L. Van de Verg, D. Hoover and R. L. Warren, “Cloning of a Brucella melitensis Group 3 Antigen Gene Encoding Omp28, a Protein Recognized by the Humoral Immune Response during Human Brucellosis,” Infection and Immunity, Vol. 64, No. 7, 1996, pp. 2490-2499.
[48] R. M. Roop 2nd, T. W. Fletcher, N. M. Sriranganathan, S. M. Boyle and G. G. Schurig, “Identification of an Immunoreactive Brucella abortus HtrA Stress Response Protein Homolog,” Infection and Immunity, Vol. 62, No. 3, 1994, pp. 1000-1007.
[49] A. Tibor, V. Weynants, P. Denoel, B. Lichtfouse, X. De Bolle, E. Saman, J. N. Limet and J. J. Letesson, “Molecular Cloning, Nucleotide Sequence, and Occurrence of a 16.5-Kilodalton Outer Membrane Protein of Brucella abortus with Similarity to Pal Lipoproteins,” Infection and Immunity, Vol. 62, No. 9, 1994, pp. 3633-3639.
[50] J. P. Connolly, D. Comerci, T. G. Alefantis, A. Walz, M. Quan, R. Chafin, P. Grewal, C. V. Mujer, R. A. Ugalde and V. G. DelVecchio, “Proteomic Analysis of Brucella abortus Cell Envelope and Identification of Immunogenic Candidate Proteins for Vaccine Development,” Proteomics, Vol. 6, No. 13, 2006, pp. 3767-3780. doi:10.1002/pmic.200500730
[51] W. Tan , X. R. Wang, Y. Nie, C. Wang, L. Q. Cheng, X. C. Wang, R. Zhang and G. M. Yan, “Recombinant VirB5 Protein as a Potential Serological Marker for the Diagnosis of Bovine Brucellosis,” Molecular and Cellular Probes, Vol. 26, No. 3, 2012, pp. 127-131. doi:10.1016/j.mcp.2012.02.003
[52] H. G. Rolarn, A. B. den Hartigh, M. Kahl-McDonagh, T. Ficht, L. G. Adams and R. M. Tsolis, “VirB12 Is a Serological Marker of Brucella Infection in Experimental and Natural Hosts,” Clinical and Vaccine Immunology, Vol. 15, No. 2, 2008, pp. 208-214. doi:10.1128/CVI.00374-07
[53] A. P. Cannella, R. M. Tsolis, L. Liang, P. L. Felgner, M. Saito, A. Sette, E. Gotuzzo and J. M. Vinetz, “AntigenSpecific Acquired Immunity in Human Brucellosis: Implications for Diagnosis, Prognosis, and Vaccine Development,” Frontiers in Cellular and Infection Microbiology, Vol. 2, 2012, p. 1. doi:10.3389/fcimb.2012.00001
[54] X. Yang, M. Hudson, N. Walters, R. F. Bargatze and D. W. Pascual, “Selection of Protective Epitopes for Brucella melitensis by DNA Vaccination,” Infection and Immunity, Vol. 73, No. 11, 2005, pp. 7297-7303. doi:10.1128/IAI.73.11.7297-7303.2005
[55] L. Liang, D. Leng, C. Burk, R. Nakajima-Sasaki, M. A. Kayala, V. L. Atluri, J. Pablo, B. Unal, T. A. Ficht, E. Gotuzzo, M. Saito, W. J. Morrow, X. Liang, P. Baldi, R. H. Gilman, J. M. Vinetz, R. M. Tsolis and P. L. Felgner, “Large Scale Immune Profiling of Infected Humans and Goats Reveals Differential Recognition of Brucella melitensis Antigens,” PLOS Neglected Tropical Diseases, Vol. 4, No. 5, 2010, p. e673. doi:10.1371/journal.pntd.0000673
[56] P. S. Chain, D. J. Comerci, M. E. Tolmasky, F. W. Larimer, S. A. Malfatti, L. M. Vergez, F. Aguero, M. L. Land, R. A. Ugalde and E. Garcia, “Whole-Genome Analyses of Speciation Events in Pathogenic Brucellae,” Infection and Immunity, Vol. 73, No. 12, 2005, pp. 8353-8361. doi:10.1128/IAI.73.12.8353-8361.2005
[57] A. Dricot, J. F. Rual, P. Lamesch, N. Bertin, D. Dupuy, T. Hao, C. Lambert, R. Hallez, J. M. Delroisse, J. Vandenhaute, I. Lopez-Goni, I. Moriyon, J. M. Garcia-Lobo, F. J. Sangari, A. P. Macmillan, S. J. Cutler, A. M. Whatmore, S. Bozak, R. Sequerra, L. Doucette-Stamm, M. Vidal, D. E. Hill, J. J. Letesson and X. De Bolle, “Generation of the Brucella melitensis ORFeome Version 1.1,” Genome Research, Vol. 14, No. 10B, 2004, pp. 2201-2206. doi:10.1101/gr.2456204
[58] C. Viadas, M. C. Rodriguez, J. M. Garcia-Lobo, F. J. Sangari and I. Lopez-Goni, “Construction and Evaluation of an ORFeome-Based Brucella Whole-Genome DNA Microarray,” Microbial Pathogenesis, Vol. 47, No. 4, 2009, pp. 189-195. doi:10.1016/j.micpath.2009.06.002
[59] L. Liang, X. Tan, S. Juarez, H. Villaverde, J. Pablo, R. Nakajima-Sasaki, E. Gotuzzo, M. Saito, G. Hermanson, D. Molina, S. Felgner, W. J. Morrow, X. Liang, R. H. Gilman, D. H. Davies, R. M. Tsolis, J. M. Vinetz and P. L. Felgner, “Systems Biology Approach Predicts Antibody Signature Associated with Brucella melitensis Infection in Humans,” Journal of Proteome Research, Vol. 10, No. 10, 2011, pp. 4813-4824. doi:10.1021/pr200619r
[60] A. P. Cannella, J. C. Lin, L. Liang, V. Atluri, E. Gotuzzo, P. L. Felgner, R. M. Tsolis and J. M. Vinetz, “Serial Kinetics of the Antibody Response against the Complete Brucella melitensis ORFeome in Focal Vertebral Brucellosis,” Journal of Clinical Microbiology, Vol. 50, No. 3, 2012, pp. 922-926. doi:10.1128/JCM.05298-11
[61] Y. He, “Analyses of Brucella Pathogenesis, Host Immunity, and Vaccine Targets Using Systems Biology and Bioinformatics,” Frontiers in Cellular and Infection Microbiology, Vol. 2, 2012, p. 2. doi:10.3389/fcimb.2012.00002
[62] L. Ferreira, S. V. Castano, F. Sanchez-Juanes, S. Gonzalez-Cabrero, F. Menegotto, A. Orduna-Domingo, J. M. Gonzales-Buitrago and J. L. Munoz-Bellido, “Identification of Brucella by MALDI-TOF Mass Spectrometry. Fast and Reliable Identification from Agar Plates and Blood Cultures,” PLoS One, Vol. 5, No. 12, 2010, pp. 1-8. doi:10.1371/journal.pone.0014235

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