Jundishapur Journal of Microbiology

Published by: Kowsar

Laboratory Diagnostic Procedures for Human Brucellosis: An Overview of Existing Approaches

Afshar Etemadi ORCID 1 , Rezvan Moniri ORCID 2 , * , Heinrich Neubauer ORCID 3 , Yasaman Dasteh Goli ORCID 4 and Saeed Alamian ORCID 5
Authors Information
1 Kashan University of Medical Sciences, Kashan, Iran
2 Department of Microbiology, Faculty of Medicine, Kashan University of Medical Sciences, Kashan, Iran
3 Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Bacterial Infections and Zoonoses, Jena, Germany
4 University of Maryland College Park, Maryland, United States
5 Brucellosis Department, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran
Article information
  • Jundishapur Journal of Microbiology: 12 (5); e91200
  • Published Online: May 20, 2019
  • Article Type: Review Article
  • Received: March 3, 2019
  • Revised: April 4, 2019
  • Accepted: April 16, 2019
  • DOI: 10.5812/jjm.91200

To Cite: Etemadi A , Moniri R, Neubauer H , Dasteh Goli Y, Alamian S . Laboratory Diagnostic Procedures for Human Brucellosis: An Overview of Existing Approaches, Jundishapur J Microbiol. Online ahead of Print ; 12(5):e91200. doi: 10.5812/jjm.91200.

Copyright © 2019, Author(s). This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.
1. Context
2. Evidence Acquisition
3. Results
4. Conclusions
  • 1. Pappas G, Papadimitriou P, Akritidis N, Christou L, Tsianos EV. The new global map of human brucellosis. Lancet Infect Dis. 2006;6(2):91-9. doi: 10.1016/S1473-3099(06)70382-6. [PubMed: 16439329].
  • 2. Lucero NE, Ayala SM, Escobar GI, Jacob NR. Brucella isolated in humans and animals in Latin America from 1968 to 2006. Epidemiol Infect. 2008;136(4):496-503. doi: 10.1017/S0950268807008795. [PubMed: 17559694]. [PubMed Central: PMC2870831].
  • 3. Wolfram JH, Butaev MK, Duysheev A, Gabbasova AR, Khasanov OS, Kulakov YK, et al. Epidemiology chapter. Vaccine. 2010;28 Suppl 5:F77-84. doi: 10.1016/j.vaccine.2010.04.050. [PubMed: 20850689].
  • 4. Alavi SM, Alavi L. Treatment of brucellosis: A systematic review of studies in recent twenty years. Caspian J Intern Med. 2013;4(2):636-41. [PubMed: 24009951]. [PubMed Central: PMC3755828].
  • 5. Demirturk N, Demirdal T, Erben N, Demir S, Asci Z, Kilit TP, et al. Brucellosis: A retrospective evaluation of 99 cases and review of brucellosis treatment. Trop Doct. 2008;38(1):59-62. doi: 10.1258/td.2006.006266. [PubMed: 18302876].
  • 6. Buzgan T, Karahocagil MK, Irmak H, Baran AI, Karsen H, Evirgen O, et al. Clinical manifestations and complications in 1028 cases of brucellosis: A retrospective evaluation and review of the literature. Int J Infect Dis. 2010;14(6):e469-78. doi: 10.1016/j.ijid.2009.06.031. [PubMed: 19910232].
  • 7. Ariza J, Pellicer T, Pallares R, Foz A, Gudiol F. Specific antibody profile in human brucellosis. Clin Infect Dis. 1992;14(1):131-40. doi: 10.1093/clinids/14.1.131. [PubMed: 1571417].
  • 8. Franc KA, Krecek RC, Hasler BN, Arenas-Gamboa AM. Brucellosis remains a neglected disease in the developing world: A call for interdisciplinary action. BMC Public Health. 2018;18(1):125. doi: 10.1186/s12889-017-5016-y. [PubMed: 29325516]. [PubMed Central: PMC5765637].
  • 9. Gupte S, Kaur T. Diagnostic approach to brucellosis. J Trop Dis. 2016;4(1). doi: 10.4172/2329-891X.1000e109.
  • 10. Al Dahouk S, Tomaso H, Nockler K, Neubauer H, Frangoulidis D. Laboratory-based diagnosis of brucellosis--A review of the literature. Part I: Techniques for direct detection and identification of Brucella spp. Clin Lab. 2003;49(9-10):487-505. [PubMed: 14572205].
  • 11. Hadush A, Pal M. Brucellosis - An infectious re-emerging bacterial zoonosis of global importance. Int J Livestock Res. 2013;3(1):28. doi: 10.5455/ijlr.20130305064802.
  • 12. Christopher S, Umapathy BL, Ravikumar KL. Brucellosis: Review on the recent trends in pathogenicity and laboratory diagnosis. J Lab Physicians. 2010;2(2):55-60. doi: 10.4103/0974-2727.72149. [PubMed: 21346896]. [PubMed Central: PMC3040083].
  • 13. Singh K. Laboratory-acquired infections. Clin Infect Dis. 2009;49(1):142-7. doi: 10.1086/599104. [PubMed: 19480580].
  • 14. Erdem H, Kilic S, Sener B, Acikel C, Alp E, Karahocagil M, et al. Diagnosis of chronic brucellar meningitis and meningoencephalitis: The results of the Istanbul-2 study. Clin Microbiol Infect. 2013;19(2):E80-6. doi: 10.1111/1469-0691.12092. [PubMed: 23210984].
  • 15. Yagupsky P. Detection of Brucellae in blood cultures. J Clin Microbiol. 1999;37(11):3437-42. [PubMed: 10523530]. [PubMed Central: PMC85661].
  • 16. Ruiz J, Lorente I, Perez J, Simarro E, Martinez-Campos L. Diagnosis of brucellosis by using blood cultures. J Clin Microbiol. 1997;35(9):2417-8. [PubMed: 9276429]. [PubMed Central: PMC229981].
  • 17. Espinosa BJ, Chacaltana J, Mulder M, Franco MP, Blazes DL, Gilman RH, et al. Comparison of culture techniques at different stages of brucellosis. Am J Trop Med Hyg. 2009;80(4):625-7. doi: 10.4269/ajtmh.2009.80.625. [PubMed: 19346389].
  • 18. Mangalgi S, Sajjan A. Comparison of three blood culture techniques in the diagnosis of human brucellosis. J Lab Physicians. 2014;6(1):14-7. doi: 10.4103/0974-2727.129084. [PubMed: 24696554]. [PubMed Central: PMC3969635].
  • 19. Akpinar O. Historical perspective of brucellosis: A microbiological and epidemiological overview. Infez Med. 2016;24(1):77-86. [PubMed: 27031903].
  • 20. Al Dahouk S, Scholz HC, Tomaso H, Bahn P, Gollner C, Karges W, et al. Differential phenotyping of Brucella species using a newly developed semi-automated metabolic system. BMC Microbiol. 2010;10:269. doi: 10.1186/1471-2180-10-269. [PubMed: 20969797]. [PubMed Central: PMC2984481].
  • 21. Ciocchini AE, Rey Serantes DA, Melli LJ, Iwashkiw JA, Deodato B, Wallach J, et al. Development and validation of a novel diagnostic test for human brucellosis using a glyco-engineered antigen coupled to magnetic beads. PLoS Negl Trop Dis. 2013;7(2). e2048. doi: 10.1371/journal.pntd.0002048. [PubMed: 23459192]. [PubMed Central: PMC3573069].
  • 22. Smits HL, Abdoel TH, Solera J, Clavijo E, Diaz R. Immunochromatographic Brucella-specific immunoglobulin M and G lateral flow assays for rapid serodiagnosis of human brucellosis. Clin Diagn Lab Immunol. 2003;10(6):1141-6. doi: 10.1128/CDLI.10.6.1141-1146.2003. [PubMed: 14607880]. [PubMed Central: PMC262433].
  • 23. Fadeel MA, Hoffmaster AR, Shi J, Pimentel G, Stoddard RA. Comparison of four commercial IgM and IgG ELISA kits for diagnosing brucellosis. J Med Microbiol. 2011;60(Pt 12):1767-73. doi: 10.1099/jmm.0.033381-0. [PubMed: 21835974].
  • 24. Solis Garcia Del Pozo J, Lorente Ortuno S, Navarro E, Solera J. Detection of IgM antibrucella antibody in the absence of IgGs: A challenge for the clinical interpretation of brucella serology. PLoS Negl Trop Dis. 2014;8(12). e3390. doi: 10.1371/journal.pntd.0003390. [PubMed: 25474572]. [PubMed Central: PMC4256177].
  • 25. Casanova A, Ariza J, Rubio M, Masuet C, Diaz R. BrucellaCapt versus classical tests in the serological diagnosis and management of human brucellosis. Clin Vaccine Immunol. 2009;16(6):844-51. doi: 10.1128/CVI.00348-08. [PubMed: 19369480]. [PubMed Central: PMC2691052].
  • 26. Xu J, Qiu Y, Cui M, Ke Y, Zhen Q, Yuan X, et al. Sustained and differential antibody responses to virulence proteins of Brucella melitensis during acute and chronic infections in human brucellosis. Eur J Clin Microbiol Infect Dis. 2013;32(3):437-47. doi: 10.1007/s10096-012-1767-7. [PubMed: 23224716].
  • 27. Lucero NE, Escobar GI, Ayala SM, Silva Paulo P, Nielsen K. Fluorescence polarization assay for diagnosis of human brucellosis. J Med Microbiol. 2003;52(Pt 10):883-7. doi: 10.1099/jmm.0.05217-0. [PubMed: 12972582].
  • 28. Nielsen K, Gall D. Fluorescence polarization assay for the diagnosis of brucellosis: A review. J Immunoassay Immunochem. 2001;22(3):183-201. doi: 10.1081/IAS-100104705. [PubMed: 11506271].
  • 29. Gupte S, Kaur T. Diagnosis of human Brucellosis. J Trop Dis. 2016;4(1). doi: 10.4172/2329-891x.1000185.
  • 30. Lin F, Xu Y, Chang Y, Liu C, Jia X, Ling B. Molecular characterization of reduced susceptibility to biocides in clinical isolates of Acinetobacter baumannii. Front Microbiol. 2017;8:1836. doi: 10.3389/fmicb.2017.01836. [PubMed: 29018420]. [PubMed Central: PMC5622949].
  • 31. Colmenero JD, Morata P, Ruiz-Mesa JD, Bautista D, Bermudez P, Bravo MJ, et al. Multiplex real-time polymerase chain reaction: A practical approach for rapid diagnosis of tuberculous and brucellar vertebral osteomyelitis. Spine (Phila Pa 1976). 2010;35(24):E1392-6. doi: 10.1097/BRS.0b013e3181e8eeaf. [PubMed: 21030888].
  • 32. Queipo-Ortuno MI, Colmenero JD, Bermudez P, Bravo MJ, Morata P. Rapid differential diagnosis between extrapulmonary tuberculosis and focal complications of brucellosis using a multiplex real-time PCR assay. PLoS One. 2009;4(2). e4526. doi: 10.1371/journal.pone.0004526. [PubMed: 19225565]. [PubMed Central: PMC2639699].
  • 33. Al Dahouk S, Nockler K. Implications of laboratory diagnosis on brucellosis therapy. Expert Rev Anti Infect Ther. 2011;9(7):833-45. doi: 10.1586/eri.11.55. [PubMed: 21810055].
  • 34. Navarro E, Escribano J, Fernandez J, Solera J. Comparison of three different PCR methods for detection of Brucella spp in human blood samples. FEMS Immunol Med Microbiol. 2002;34(2):147-51. doi: 10.1111/j.1574-695X.2002.tb00616.x. [PubMed: 12381466].
  • 35. Alamian S, Esmaelizad M, Zahraei T, Etemadi A, Mohammadi M, Afshar D, et al. A novel PCR assay for detecting Brucella abortus and Brucella melitensis. Osong Public Health Res Perspect. 2017;8(1):65-70. doi: 10.24171/j.phrp.2017.8.1.09. [PubMed: 28443226]. [PubMed Central: PMC5402848].
  • 36. Etemady A, Mohammdi M, Esmaelizad M, Alamian S, Vahedi F, Aghaeipour K, et al. Genetic characterization of the wboA gene from the predominant biovars of Brucella isolates in Iran. Electron Physician. 2015;7(6):1381-6. doi: 10.14661/1381. [PubMed: 26516446]. [PubMed Central: PMC4623799].
  • 37. Yu WL, Nielsen K. Review of detection of Brucella spp. by polymerase chain reaction. Croat Med J. 2010;51(4):306-13. doi: 10.3325/cmj.2010.51.306. [PubMed: 20718083]. [PubMed Central: PMC2931435].
  • 38. Baddour MM, Alkhalifa DH. Evaluation of three polymerase chain reaction techniques for detection of Brucella DNA in peripheral human blood. Can J Microbiol. 2008;54(5):352-7. doi: 10.1139/w08-017. [PubMed: 18449219].
  • 39. Mitka S, Anetakis C, Souliou E, Diza E, Kansouzidou A. Evaluation of different PCR assays for early detection of acute and relapsing brucellosis in humans in comparison with conventional methods. J Clin Microbiol. 2007;45(4):1211-8. doi: 10.1128/JCM.00010-06. [PubMed: 17267626]. [PubMed Central: PMC1865811].
  • 40. Wang Y, Wang Z, Zhang Y, Bai L, Zhao Y, Liu C, et al. Polymerase chain reaction-based assays for the diagnosis of human brucellosis. Ann Clin Microbiol Antimicrob. 2014;13:31. doi: 10.1186/s12941-014-0031-7. [PubMed: 25082566]. [PubMed Central: PMC4236518].
  • 41. Imaoka K, Kimura M, Suzuki M, Kamiyama T, Yamada A. Simultaneous detection of the genus Brucella by combinatorial PCR. Jpn J Infect Dis. 2007;60(2-3):137-9. [PubMed: 17515651].
  • 42. Mayer-Scholl A, Draeger A, Gollner C, Scholz HC, Nockler K. Advancement of a multiplex PCR for the differentiation of all currently described Brucella species. J Microbiol Methods. 2010;80(1):112-4. doi: 10.1016/j.mimet.2009.10.015. [PubMed: 19887090].
  • 43. Kumar S, Tuteja U, Sarika K, Singh D, Kumar A, Kumar O. Rapid multiplex PCR assay for the simultaneous detection of the Brucella Genus, B. abortus, B. melitensis, and B. suis. J Microbiol Biotechnol. 2011;21(1):89-92. doi: 10.4014/jmb.1007.07051. [PubMed: 21301197].
  • 44. Kamal IH, Al Gashgari B, Moselhy SS, Kumosani TA, Abulnaja KO. Two-stage PCR assay for detection of human brucellosis in endemic areas. BMC Infect Dis. 2013;13:145. doi: 10.1186/1471-2334-13-145. [PubMed: 23517532]. [PubMed Central: PMC3614503].
  • 45. Garofolo G, Ancora M, Di Giannatale E. MLVA-16 loci panel on Brucella spp. using multiplex PCR and multicolor capillary electrophoresis. J Microbiol Methods. 2013;92(2):103-7. doi: 10.1016/j.mimet.2012.11.007. [PubMed: 23174277].
  • 46. Mirnejad R, Mohamadi M, Piranfar V, Mortazavi SM, Kachuei R. A duplex PCR for rapid and simultaneous detection of Brucella spp. in human blood samples. Asian Pac J Trop Med. 2013;6(6):453-6. doi: 10.1016/S1995-7645(13)60073-5. [PubMed: 23711705].
  • 47. Al-Nakkas A, Mustafa AS, Wright SG. Large-scale evaluation of a single-tube nested PCR for the laboratory diagnosis of human brucellosis in Kuwait. J Med Microbiol. 2005;54(Pt 8):727-30. doi: 10.1099/jmm.0.45772-0. [PubMed: 16014425].
  • 48. Hasanjani Roushan MR, Marashi SM, Moulana Z. Polymerase chain reaction-based assays for the diagnosis of active and relapsed cases of human brucellosis. Am J Trop Med Hyg. 2016;95(6):1272-6. doi: 10.4269/ajtmh.16-0344. [PubMed: 27928078]. [PubMed Central: PMC5154438].
  • 49. Cirak MY, Hizel K. [The value of polymerase chain reaction methods targeting two different gene regions for the diagnosis of brucellosis]. Mikrobiyol Bul. 2002;36(3-4):271-6. Turkish. [PubMed: 12838660].
  • 50. Lin GZ, Zheng FY, Zhou JZ, Gong XW, Wang GH, Cao XA, et al. Loop-mediated isothermal amplification assay targeting the omp25 gene for rapid detection of Brucella spp. Mol Cell Probes. 2011;25(2-3):126-9. doi: 10.1016/j.mcp.2011.01.001. [PubMed: 21232598].
  • 51. Fekete A, Bantle JA, Halling SM, Stich RW. Amplification fragment length polymorphism in Brucella strains by use of polymerase chain reaction with arbitrary primers. J Bacteriol. 1992;174(23):7778-83. doi: 10.1128/jb.174.23.7778-7783.1992. [PubMed: 1360006]. [PubMed Central: PMC207493].
  • 52. Al Dahouk S, Tomaso H, Prenger-Berninghoff E, Splettstoesser WD, Scholz HC, Neubauer H. Identification of brucella species and biotypes using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Crit Rev Microbiol. 2005;31(4):191-6. doi: 10.1080/10408410500304041. [PubMed: 16417200].
  • 53. Vrioni G, Gartzonika C, Kostoula A, Boboyianni C, Papadopoulou C, Levidiotou S. Application of a polymerase chain reaction enzyme immunoassay in peripheral whole blood and serum specimens for diagnosis of acute human brucellosis. Eur J Clin Microbiol Infect Dis. 2004;23(3):194-9. doi: 10.1007/s10096-003-1082-4. [PubMed: 14986157].
  • 54. Garibyan L, Avashia N. Polymerase chain reaction. J Invest Dermatol. 2013;133(3):1-4. doi: 10.1038/jid.2013.1. [PubMed: 23399825]. [PubMed Central: PMC4102308].
  • 55. Kattar MM, Zalloua PA, Araj GF, Samaha-Kfoury J, Shbaklo H, Kanj SS, et al. Development and evaluation of real-time polymerase chain reaction assays on whole blood and paraffin-embedded tissues for rapid diagnosis of human brucellosis. Diagn Microbiol Infect Dis. 2007;59(1):23-32. doi: 10.1016/j.diagmicrobio.2007.04.002. [PubMed: 17532591].
  • 56. Debeaumont C, Falconnet PA, Maurin M. Real-time PCR for detection of Brucella spp. DNA in human serum samples. Eur J Clin Microbiol Infect Dis. 2005;24(12):842-5. doi: 10.1007/s10096-005-0064-0. [PubMed: 16341519].
  • 57. Queipo-Ortuno MI, Colmenero JD, Bravo MJ, Garcia-Ordonez MA, Morata P. Usefulness of a quantitative real-time PCR assay using serum samples to discriminate between inactive, serologically positive and active human brucellosis. Clin Microbiol Infect. 2008;14(12):1128-34. doi: 10.1111/j.1469-0691.2008.02095.x. [PubMed: 19046166].
  • 58. Zhang B, Wear DJ, Stojadinovic A, Izadjoo M. Sequential real-time PCR assays applied to identification of genomic signatures in formalin-fixed paraffin-embedded tissues: A case report about brucella-induced osteomyelitis. Mil Med. 2013;178(1):88-94. doi: 10.7205/MILMED-D-12-00274. [PubMed: 23356125].
  • 59. Colmenero JD, Clavijo E, Morata P, Bravo MJ, Queipo-Ortuno MI. Quantitative real-time polymerase chain reaction improves conventional microbiological diagnosis in an outbreak of brucellosis due to ingestion of unpasteurized goat cheese. Diagn Microbiol Infect Dis. 2011;71(3):294-6. doi: 10.1016/j.diagmicrobio.2011.06.016. [PubMed: 21855249].
  • 60. Bounaadja L, Albert D, Chenais B, Henault S, Zygmunt MS, Poliak S, et al. Real-time PCR for identification of Brucella spp.: A comparative study of IS711, bcsp31 and per target genes. Vet Microbiol. 2009;137(1-2):156-64. doi: 10.1016/j.vetmic.2008.12.023. [PubMed: 19200666].
  • 61. Sanjuan-Jimenez R, Colmenero JD, Bermudez P, Alonso A, Morata P. Amplicon DNA melting analysis for the simultaneous detection of Brucella spp and Mycobacterium tuberculosis complex. Potential use in rapid differential diagnosis between extrapulmonary tuberculosis and focal complications of brucellosis. PLoS One. 2013;8(3). e58353. doi: 10.1371/journal.pone.0058353. [PubMed: 23520501]. [PubMed Central: PMC3592798].
  • 62. Gopaul KK, Sells J, Bricker BJ, Crasta OR, Whatmore AM. Rapid and reliable single nucleotide polymorphism-based differentiation of Brucella live vaccine strains from field strains. J Clin Microbiol. 2010;48(4):1461-4. doi: 10.1128/JCM.02193-09. [PubMed: 20181906]. [PubMed Central: PMC2849582].
  • 63. Foster JT, Okinaka RT, Svensson R, Shaw K, De BK, Robison RA, et al. Real-time PCR assays of single-nucleotide polymorphisms defining the major Brucella clades. J Clin Microbiol. 2008;46(1):296-301. doi: 10.1128/JCM.01496-07. [PubMed: 18032628]. [PubMed Central: PMC2224295].
  • 64. Maio E, Begeman L, Bisselink Y, van Tulden P, Wiersma L, Hiemstra S, et al. Identification and typing of Brucella spp. in stranded harbour porpoises (Phocoena phocoena) on the Dutch coast. Vet Microbiol. 2014;173(1-2):118-24. doi: 10.1016/j.vetmic.2014.07.010. [PubMed: 25115787].
  • 65. Seng P, Drancourt M, Gouriet F, La Scola B, Fournier PE, Rolain JM, et al. Ongoing revolution in bacteriology: Routine identification of bacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Clin Infect Dis. 2009;49(4):543-51. doi: 10.1086/600885. [PubMed: 19583519].
  • 66. Ferreira L, Vega Castano S, Sanchez-Juanes F, Gonzalez-Cabrero S, Menegotto F, Orduna-Domingo A, et al. Identification of Brucella by MALDI-TOF mass spectrometry. Fast and reliable identification from agar plates and blood cultures. PLoS One. 2010;5(12). e14235. doi: 10.1371/journal.pone.0014235. [PubMed: 21151913]. [PubMed Central: PMC2997794].
  • 67. Mesureur J, Ranaldi S, Monnin V, Girard V, Arend S, Welker M, et al. A simple and safe protocol for preparing brucella samples for matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis. J Clin Microbiol. 2016;54(2):449-52. doi: 10.1128/JCM.02730-15. [PubMed: 26582837]. [PubMed Central: PMC4733191].
  • 68. Wareth G, Eravci M, Weise C, Roesler U, Melzer F, Sprague LD, et al. Comprehensive identification of immunodominant proteins of Brucella abortus and Brucella melitensis using antibodies in the sera from naturally infected hosts. Int J Mol Sci. 2016;17(5). doi: 10.3390/ijms17050659. [PubMed: 27144565]. [PubMed Central: PMC4881485].
  • 69. Kim JY, Sung SR, Lee K, Lee HK, Kang SI, Lee JJ, et al. Immunoproteomics of Brucella abortus RB51 as candidate antigens in serological diagnosis of brucellosis. Vet Immunol Immunopathol. 2014;160(3-4):218-24. doi: 10.1016/j.vetimm.2014.05.009. [PubMed: 24908638].
  • 70. Pajuaba AC, Silva DA, Almeida KC, Cunha-Junior JP, Pirovani CP, Camillo LR, et al. Immunoproteomics of Brucella abortus reveals differential antibody profiles between S19-vaccinated and naturally infected cattle. Proteomics. 2012;12(6):820-31. doi: 10.1002/pmic.201100185. [PubMed: 22539433].

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