{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,11]],"date-time":"2026-10-11T17:25:33Z","timestamp":1791739533050,"version":"4.3.4"},"reference-count":94,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2021,12,14]],"date-time":"2021-12-14T00:00:00Z","timestamp":1639440000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Brucella spp. are Gram-negative, non-motile, non-spore-forming, slow-growing, facultative intracellular bacteria causing brucellosis. Brucellosis is an endemic of specific geographic areas and, although underreported, represents the most common zoonotic infection, with an annual global incidence of 500,000 cases among humans. Humans represent an occasional host where the infection is mainly caused by B. melitensis, which is the most virulent; B. abortus; B. suis; and B. canis. A microbiological analysis is crucial to identifying human cases because clinical symptoms of human brucellosis are variable and aspecific. The laboratory diagnosis is based on three different microbiological approaches: (i) direct diagnosis by culture, (ii) indirect diagnosis by serological tests, and (iii) direct rapid diagnosis by molecular PCR-based methods. Despite the established experience with serological tests and highly sensitive nucleic acid amplification tests (NAATs), a culture is still considered the \u201cgold standard\u201d in the laboratory diagnosis of brucellosis due to its clinical and epidemiological relevance. Moreover, the automated BC systems now available have increased the sensitivity of BCs and shortened the time to detection of Brucella species. The main limitations of serological tests are the lack of common interpretative criteria, the suboptimal specificity due to interspecies cross-reactivity, and the low sensitivity during the early stage of disease. Despite that, serological tests remain the main diagnostic tool, especially in endemic areas because they are inexpensive, user friendly, and have high negative predictive value. Promising serological tests based on new synthetic antigens have been recently developed together with novel point-of-care tests without the need for dedicated equipment and expertise. NAATs are rapid tests that can help diagnose brucellosis in a few hours with high sensitivity and specificity. Nevertheless, the interpretation of NAAT-positive results requires attention because it may not necessarily indicate an active infection but rather a low bacterial inoculum, DNA from dead bacteria, or a patient that has recovered. Refined NAATs should be developed, and their performances should be compared with those of commercial and home-made molecular tests before being commercialized for the diagnosis of brucellosis. Here, we review and report the most common and updated microbiological diagnostic methods currently available for the laboratory diagnosis of brucellosis.<\/jats:p>","DOI":"10.3390\/pathogens10121623","type":"journal-article","created":{"date-parts":[[2021,12,14]],"date-time":"2021-12-14T22:06:10Z","timestamp":1639519570000},"page":"1623","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":168,"title":["Microbiological Laboratory Diagnosis of Human Brucellosis: An Overview"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5850-2810","authenticated-orcid":false,"given":"Giovanni","family":"Di Bonaventura","sequence":"first","affiliation":[{"name":"Department of Medical, Oral and Biotechnological Sciences, \u201cG. d\u2019Annunzio\u201d University of Chieti-Pescara, Via Luigi Polacchi 11, 66100 Chieti, Italy"},{"name":"Center for Advanced Studies and Technology (CAST), \u201cG. d\u2019Annunzio\u201d University of Chieti-Pescara, Via Luigi Polacchi 11, 66100 Chieti, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia","family":"Angeletti","sequence":"additional","affiliation":[{"name":"Department of Medicine, Campus Biomedico University, Via Alvaro del Portillo 200, 00128 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"Ianni","sequence":"additional","affiliation":[{"name":"Department of Medicine, Campus Biomedico University, Via Alvaro del Portillo 200, 00128 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5585-9475","authenticated-orcid":false,"given":"Tommasangelo","family":"Petitti","sequence":"additional","affiliation":[{"name":"Department of Medicine, Campus Biomedico University, Via Alvaro del Portillo 200, 00128 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giovanni","family":"Gherardi","sequence":"additional","affiliation":[{"name":"Department of Medicine, Campus Biomedico University, Via Alvaro del Portillo 200, 00128 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,14]]},"reference":[{"key":"ref_1","unstructured":"Araj, G.F. (2019). Brucella. Manual for Clinical Microbiology, ASM Press. [12th ed.]."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Jiao, H., Zhou, Z., Li, B., Xiao, Y., Li, M., Zeng, H., Guo, X., and Gu, G. (2021). The mechanism of facultative intracellular parasitism of Brucella. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22073673"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Whatmore, A.M., Koylass, M.S., Muchowski, J., Edwards-Smallbone, J., Gopaul, K.K., and Perrett, L.L. (2016). Extended multilocus sequence analysis to describe the global population structure of the genus Brucella: Phylogeography and relationship to biovars. Front. Microbiol., 7.","DOI":"10.3389\/fmicb.2016.02049"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Al Dahouk, S., Scholz, H.C., Tomaso, H., Bahn, P., Gollner, C., Karges, W., Appel, B., Hensel, A., Neubauer, H., and Nockler, K. (2010). Differential phenotyping of Brucella species using a newly developed semi-automated metabolic system. BMC Microbiol., 10.","DOI":"10.1186\/1471-2180-10-269"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e135","DOI":"10.1086\/523726","article-title":"The many faces of human-to-human transmission of brucellosis: Congenital infection and outbreak of nosocomial disease related to an unrecognized clinical case","volume":"45","author":"Mesner","year":"2007","journal-title":"Clin. Infect. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"775","DOI":"10.1016\/S1473-3099(07)70286-4","article-title":"Human brucellosis","volume":"7","author":"Franco","year":"2007","journal-title":"Lancet Infect. Dis."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1186\/s41182-020-00272-1","article-title":"One health approach to tackle brucellosis: A systematic review","volume":"48","author":"Ghanbari","year":"2020","journal-title":"Trop. Med. Health"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"812","DOI":"10.1007\/s15010-021-01586-w","article-title":"The current therapeutical strategies in human brucellosis","volume":"49","author":"Bosilkovski","year":"2021","journal-title":"Infection"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2325","DOI":"10.1056\/NEJMra050570","article-title":"Brucellosis","volume":"352","author":"Pappas","year":"2005","journal-title":"N. Engl. J. Med."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Yagupsky, P., Morata, P., and Colmenero, J.D. (2019). Laboratory diagnosis of human brucellosis. Clin. Microbiol. Rev., 33.","DOI":"10.1128\/CMR.00073-19"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"495","DOI":"10.2460\/ajvr.78.4.495","article-title":"Evaluation of a hypervariable octameric oligonucleotide fingerprints assay for identification of and discrimination between wild-type and vaccine strains of Brucella melitensis","volume":"78","author":"Tang","year":"2017","journal-title":"Am. J. Vet. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1599","DOI":"10.1089\/vbz.2011.0704","article-title":"Limitations of the standard agglutination test for detecting patients with Brucella melitensis bacteremia","volume":"11","author":"Shemesh","year":"2011","journal-title":"Vector Borne Zoonotic Dis."},{"key":"ref_13","unstructured":"Baron, E.J. (2005). Blood Cultures IV. Cumitech 1C, ASM Press."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"S29","DOI":"10.1016\/j.ijantimicag.2007.06.011","article-title":"Challenges in Brucella bacteremia","volume":"30","author":"Pappas","year":"2007","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1382","DOI":"10.1128\/jcm.35.6.1382-1384.1997","article-title":"Comparison of BACTEC 9240 Peds Plus medium and Isolator 1.5 Microbial ube for detection of Brucella melitensis from blood cultures","volume":"35","author":"Yagupsky","year":"1997","journal-title":"J. Clin. Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"3200","DOI":"10.1128\/jcm.31.12.3200-3203.1993","article-title":"Factors affecting detection of Brucella melitensis by BACTEC NR730, a nonradiometric system for hemocultures","volume":"31","author":"Gamazo","year":"1993","journal-title":"J. Clin. Microbiol."},{"key":"ref_17","first-page":"776","article-title":"Rapid detection of Brucella by an automated blood culture system at a tertiary care hospital of north India","volume":"139","author":"Raj","year":"2014","journal-title":"Indian J. Med. Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1038","DOI":"10.1046\/j.1469-0691.2003.00714.x","article-title":"Prospective evaluation of blood cultures in a Turkish university hospital: Epidemiology, microbiology and patient outcome","volume":"9","author":"Esel","year":"2003","journal-title":"Clin. Microbiol. Infect."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1016\/S0213-005X(01)72633-X","article-title":"Deteccion de Brucella melitensis por el sistema BACTEC 9050","volume":"19","author":"Lepe","year":"2001","journal-title":"Enferm. Infecc. Microbiol. Clin."},{"key":"ref_20","first-page":"21","article-title":"Development and evaluation of TUMS medium, a novel biphasic culture medium for isolation of Brucella spp. from patients","volume":"1","author":"Fatolahzadeh","year":"2009","journal-title":"Iran. J. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1007\/BF02447926","article-title":"Rapid detection of Brucella melitensis from blood cultures by a commercial system","volume":"16","author":"Yagupsky","year":"1997","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"942","DOI":"10.1128\/JCM.02320-16","article-title":"The Bactec FX blood culture system detects Brucella melitensis bacteremia in adult patients within the routine 1-week incubation period","volume":"55","author":"Sagi","year":"2017","journal-title":"J. Clin. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1016\/j.resmic.2005.10.005","article-title":"Evaluation of the Bactec microbial detection system for culturing miscellaneous sterile body fluids","volume":"157","author":"Akcam","year":"2006","journal-title":"Res. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1056\/NEJM195708012570508","article-title":"Isolation of Brucella abortus by percutaneous liver biopsy","volume":"257","author":"Cohen","year":"1957","journal-title":"N. Engl. J. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"204","DOI":"10.7326\/0003-4819-53-1-204","article-title":"Diagnosis of Brucella melitensis by percutaneous needle biopsy of the liver","volume":"53","author":"Fogel","year":"1960","journal-title":"Ann. Intern. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1016\/j.ijid.2007.09.004","article-title":"Bacteremia is as unpredictable as clinical manifestations in human brucellosis","volume":"12","author":"Mantur","year":"2008","journal-title":"Int. J. Infect. Dis."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1093\/infdis\/66.1.86","article-title":"Isolation of Brucella from lymph nodes","volume":"66","author":"Poston","year":"1940","journal-title":"J. Infect. Dis."},{"key":"ref_28","first-page":"73","article-title":"Laboratory diagnosis of brucellosis in man","volume":"24","year":"1961","journal-title":"Bull. World Health Org."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1093\/clinids\/21.2.283","article-title":"An overview of human brucellosis","volume":"21","author":"Young","year":"1995","journal-title":"Clin. Infect. Dis."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1007\/s10067-017-3819-y","article-title":"Sixteen Chinese pediatric brucellosis patients onset of fever in non-epidemic areas and 8 developed with osteoarticular involvement","volume":"37","author":"Wang","year":"2018","journal-title":"Clin. Rheumatol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"S12","DOI":"10.1016\/j.ijantimicag.2010.06.014","article-title":"Update on laboratory diagnosis of human brucellosis","volume":"36","author":"Araj","year":"2010","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Diaz, R., Casanova, A., Ariza, J., and Moriyon, I. (2011). The Rose Bengal Test in human brucellosis: A neglected test for the diagnosis of a neglected disease. PLoS Negl. Trop. Dis., 5.","DOI":"10.1371\/journal.pntd.0000950"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"833","DOI":"10.1586\/eri.11.55","article-title":"Implications of laboratory diagnosis on brucellosis therapy","volume":"9","author":"Nockler","year":"2011","journal-title":"Expert Rev. Anti Infect. Ther."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.phrp.2012.01.003","article-title":"Application of the microagglutination test for serologic diagnosis of human brucellosis","volume":"3","author":"Park","year":"2012","journal-title":"Osong Public Health Res. Perspect."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1128\/jcm.11.6.691-693.1980","article-title":"2-Mercaptoethanol Brucella agglutination test: Usefulness for predicting recovery from brucellosis","volume":"11","author":"Buchanan","year":"1980","journal-title":"J. Clin. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"918","DOI":"10.1093\/infdis\/157.5.918","article-title":"Specific antibodies detected during relapse of human brucellosis","volume":"157","author":"Pellicer","year":"1988","journal-title":"J. Infect. Dis."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1671","DOI":"10.7754\/Clin.Lab.2016.160120","article-title":"Comparison of a novel test (ODAK Brucella Coombs Gel Test) with commonly used serological tests in human brucellosis","volume":"62","author":"Borsa","year":"2016","journal-title":"Clin. Lab."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"108","DOI":"10.3923\/pjbs.2017.108.112","article-title":"Evaluation of a new and rapid serologic test for detecting brucellosis: Brucella Coombs gel test","volume":"20","author":"Hanci","year":"2017","journal-title":"Pak. J. Biol. Sci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1017\/S0022172400046684","article-title":"The use of the conglutinating complement fixation test in the diagnosis of human brucellosis","volume":"74","author":"Farrell","year":"1975","journal-title":"J. Hyg."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1128\/CVI.00348-08","article-title":"BrucellaCapt versus classical tests in the serological diagnosis and management of human brucellosis","volume":"16","author":"Casanova","year":"2009","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"4000","DOI":"10.1128\/JCM.38.11.4000-4005.2000","article-title":"Evaluation of an immunocapture-agglutination test (BrucellaCapt) for serodiagnosis of human brucellosis","volume":"38","author":"Orduna","year":"2000","journal-title":"J. Clin. Microbiol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"272","DOI":"10.7883\/yoken.64.272","article-title":"Evaluation of serological tests for diagnoisis of brucellosis","volume":"64","author":"Pabuccuoglu","year":"2011","journal-title":"Jpn. J. Infect. Dis."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1111\/j.1699-0463.1988.tb05286.x","article-title":"ELISA versus routine tests in the diagnosis of patients with systemic and neurobrucellosis","volume":"96","author":"Araj","year":"1988","journal-title":"APMIS"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3098","DOI":"10.1128\/JCM.00919-09","article-title":"Time resolved fluorescent resonance energy transfer assay for simple and rapid detection of anti-Brucella antibodies in ruminant serum sample","volume":"47","author":"McGiven","year":"2009","journal-title":"J. Clin. Microbiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1099\/jmm.0.05217-0","article-title":"Fluorescence polarization assay for diagnosis of human brucellosis","volume":"52","author":"Lucero","year":"2003","journal-title":"J. Med. Microbiol."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Li, G., Rong, Z., Wang, S., Zhao, H., Piao, D., Yang, X., Tian, G., and Jiang, H. (2020). Rapid detection of brucellosis using a quantum dot-based immunochromatographic test strip. PLoS Negl. Trop. Dis., 14.","DOI":"10.1371\/journal.pntd.0008557"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1128\/jcm.34.2.477-478.1996","article-title":"Rapid laboratory confirmation of human brucellosis by PCR analysis of a target sequence on the 31-kilodalton Brucella antigen DNA","volume":"34","author":"Matar","year":"1996","journal-title":"J. Clin. Microbiol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1111\/j.1574-695X.2002.tb00616.x","article-title":"Comparison of three different PCR methods for detection of Brucella spp. in human blood samples","volume":"34","author":"Navarro","year":"2002","journal-title":"FEMS Immunol. Med. Microbiol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1661","DOI":"10.1128\/JCM.39.4.1661-1664.2001","article-title":"Serum is the preferred clinical specimen for diagnosis of human brucellosis by PCR","volume":"39","author":"Zerva","year":"2001","journal-title":"J. Clin. Microbiol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"e0062","DOI":"10.1097\/MD.0000000000010062","article-title":"Real-time PCR assays for diagnosing brucellar spondylitis using formalin-fixed paraffin embedded tissues","volume":"97","author":"Li","year":"2018","journal-title":"Medicine"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"106182","DOI":"10.1016\/j.mimet.2021.106182","article-title":"Comparative study of IS711 and bcsp31-based polymerase chain reaction (PCR) for the diagnosis of human brucellosis in whole blood and serum samples","volume":"183","author":"Becker","year":"2021","journal-title":"J. Microbiol. Methods"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"3087","DOI":"10.1128\/jcm.33.12.3087-3090.1995","article-title":"Single-step PCR for detection of Brucella spp. From blood and milk of infected animals","volume":"33","year":"1995","journal-title":"J. Clin. Microbiol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.diagmicrobio.2007.04.002","article-title":"Development and evaluation of real-time polymerase chain reaction assays on whole blood and paraffin embedded tissues for rapid diagnosis of human brucellosis","volume":"59","author":"Kattar","year":"2007","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1211","DOI":"10.1128\/JCM.00010-06","article-title":"Evaluation of different PCR assays for early detection of acute and relapsing brucellosis in humans in comparison with conventional methods","volume":"45","author":"Mitka","year":"2007","journal-title":"J. Clin. Microbiol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"352","DOI":"10.1139\/W08-017","article-title":"Evaluation of three polymerase chain reaction techniques for detection of Brucella DNA in peripheral human blood","volume":"54","author":"Baddour","year":"2008","journal-title":"Can. J. Microbiol."},{"key":"ref_56","first-page":"271","article-title":"Detection of Brucella melitensis and Brucella abortus by DNA amplification","volume":"95","author":"Baily","year":"1992","journal-title":"J. Trop. Med. Hyg."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1016\/j.vetmic.2008.12.023","article-title":"Real-time PCR for identification of Brucella spp.: A comparative study of IS711, bcsp31, and per target genes","volume":"137","author":"Bounaadja","year":"2009","journal-title":"Vet. Microbiol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1128\/jcm.33.3.615-617.1995","article-title":"Specific detection of Brucella DNA by PCR","volume":"33","author":"Romero","year":"1995","journal-title":"J. Clin. Microbiol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1099\/jmm.0.45772-0","article-title":"Large-scale evaluation of a single-tube nested PCR for the laboratory diagnosis of human brucellosis in Kuwait","volume":"54","author":"Mustafa","year":"2005","journal-title":"J. Med. Microbiol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1026","DOI":"10.1038\/nbt0993-1026","article-title":"Kinetic PCR analysis: Real-time monitoring of DNA amplification reactions","volume":"11","author":"Higuchi","year":"1993","journal-title":"Nat. Biotechnol."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1016\/j.cca.2014.10.017","article-title":"Real-time PCR detection chemistry","volume":"439","author":"Navarro","year":"2015","journal-title":"Clin. Chim. Acta."},{"key":"ref_62","doi-asserted-by":"crossref","unstructured":"Sanjuan-Jimenez, R., Colmenero, J.D., Bermudez, P., Alonso, A., and Morata, P. (2013). 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, 8.","DOI":"10.1371\/journal.pone.0058353"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"130","DOI":"10.2144\/97221bi01","article-title":"Continuous fluorescence monitoring of rapid cycle DNA amplification","volume":"22","author":"Wittwer","year":"1997","journal-title":"Biotechniques"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/S0890-8508(95)80033-6","article-title":"Rapid diagnosis of Mycobacterium tuberculosis infections by ELISA-like detection polymerase chain reaction products","volume":"9","author":"Zambardi","year":"1995","journal-title":"Mol. Cell Probes"},{"key":"ref_65","doi-asserted-by":"crossref","unstructured":"Moeini-Zanjani, A., Pournajaf, A., Ferdosi-Shahandashti, E., Gholami, M., Masjedian, F., Khafri, S., and Rajabnia, R. (2020). Comparison of loop-mediated isothermal amplification and conventional PCR tests for diagnosis of common Brucella species. BMC Res. Notes, 13.","DOI":"10.1186\/s13104-020-05377-8"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"1640","DOI":"10.1128\/jcm.33.6.1640-1642.1995","article-title":"Enhancement of the Brucella AMOS PCR assay for differentiation of Brucella abortus vaccine strains S19 and RB51","volume":"33","author":"Bricker","year":"1995","journal-title":"J. Clin. Microbiol."},{"key":"ref_67","doi-asserted-by":"crossref","unstructured":"Gopaul, K.K., Sells, J., Lee, R., Beckstrom-Sternberg, S.M., Foster, J.T., and Whatmore, A.M. (2014). Development and assessment of multiplex high resolution melting assay as a tool for rapid single-tube identification of five Brucella species. BMC Res. Notes, 7.","DOI":"10.1186\/1756-0500-7-903"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"137","DOI":"10.7883\/yoken.JJID.2007.137","article-title":"Simultaneous detection of the genus Brucella by combinatorial PCR","volume":"60","author":"Imaoka","year":"2007","journal-title":"Jpn. J. Infect. Dis."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"3484","DOI":"10.1128\/JCM.00837-08","article-title":"Evaluation of a multiplex PCR assay (Bruce-ladder) for molecular typing of all Brucella species, including the vaccine strains","volume":"46","author":"Marin","year":"2008","journal-title":"J. Clin. Microbiol."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1099\/jmm.0.47507-0","article-title":"Specific detection and differentiation of Ochrobactrum anthropi, Ochrobactrum intermedium and Brucella spp. by a multiprimer PCR that targets the recA gene","volume":"57","author":"Scholz","year":"2008","journal-title":"J. Med. Microbiol."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"1828","DOI":"10.1128\/JCM.44.5.1828-1830.2006","article-title":"Rapid detection of Brucella spp. in blood cultures by fluorescence in situ hybridization","volume":"44","author":"Wellinghausen","year":"2006","journal-title":"J. Clin. Microbiol."},{"key":"ref_72","doi-asserted-by":"crossref","unstructured":"Nimri, L.F. (2003). Diagnosis of recent and relapsed cases of human brucellosis by PCR assay. BMC Infect. Dis., 3.","DOI":"10.1186\/1471-2334-3-5"},{"key":"ref_73","first-page":"15","article-title":"Evaluation of real-time PCR method for rapid diagnosis of brucellosis with different clinical manifestations","volume":"58","author":"Surucuoglu","year":"2009","journal-title":"Pol. J. Microbiol."},{"key":"ref_74","doi-asserted-by":"crossref","unstructured":"Arora, P. (2019). Molecular diagnosis of acute and chronic brucellosis in humans. Microbial Technology for the Welfare of Society. Microorganisms for Sustainability, Springer.","DOI":"10.1007\/978-981-13-8844-6"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"432","DOI":"10.3855\/jidc.7532","article-title":"Brucella suis bacteremia misidentified as Ochrobactrum anthropi by the VITEK 2 system","volume":"10","author":"Vila","year":"2016","journal-title":"J. Infect. Dev. Ctries."},{"key":"ref_76","doi-asserted-by":"crossref","unstructured":"Lista, F., Reubsaet, F.A.G., De Santis, R., Parchen, R.R., de Jong, A.L., Kieboom, J., van der Laaken, A.L., Voskamp-Visser, I.A., Fillo, S., and Jansen, H.J. (2011). Reliable identification at the species level of Brucella isolated with MALDI-TOF. BMC Microbiol., 11.","DOI":"10.1186\/1471-2180-11-267"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"3123","DOI":"10.1128\/JCM.01720-13","article-title":"Interlaboratory comparison of intact-cell matrix-assisted laser desorption ionization time of flight mass spectrometry results for identification and differentiation of Brucella spp","volume":"51","author":"Karger","year":"2013","journal-title":"J. Clin. Microbiol."},{"key":"ref_78","doi-asserted-by":"crossref","unstructured":"Mesureur, J., Arend, S., Celliere, B., Courault, P., Cotte-Pattat, P.J., Totty, H., Deol, P., Mick, V., Girard, V., and Touchberry, J. (2018). A MALDI-TOF MS database with broad genus coverage for species-level identification of Brucella. PLoS Negl. Trop. Dis., 1.","DOI":"10.1371\/journal.pntd.0006874"},{"key":"ref_79","doi-asserted-by":"crossref","unstructured":"Poonawala, H., Marrs Conner, T., and Peaper, D.R. (2018). The briefcase: Misidentification of Brucella melitensis as Ochrobactrum anthropi by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). J. Clin. Microbiol., 56.","DOI":"10.1128\/JCM.00914-17"},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"1180","DOI":"10.3201\/eid1108.041197","article-title":"Laboratory-exposures to brucellae and implications for bioterrorism","volume":"11","author":"Yagupsky","year":"2005","journal-title":"Emerg. Infect. Dis."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"1848","DOI":"10.3201\/eid1010.040076","article-title":"Laboratory-acquired brucellosis","volume":"10","author":"Noviello","year":"2004","journal-title":"Emerg. Infect. Dis."},{"key":"ref_82","unstructured":"Centers for Disease Control, National Institutes of Health (2020). Biosafety in Microbiological and Biomedical Laboratories, U.S. Department of Health and Human Services Public Health. [6th ed.]."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1093\/ije\/25.2.450","article-title":"Seroprevalence of brucellosis among high-risk people in northern Jordan","volume":"25","author":"Odeh","year":"1996","journal-title":"Int. J. Epidemiol."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1093\/clinids\/14.1.131","article-title":"Specific antibody profile in human brucellosis","volume":"14","author":"Ariza","year":"1992","journal-title":"Clin. Infect. Dis."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.medmal.2018.12.004","article-title":"Limitations of diagnostic tests for bacterial infections","volume":"49","author":"Eldin","year":"2018","journal-title":"Med. Mal. Infect."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"163","DOI":"10.20506\/rst.32.1.2205","article-title":"New developments in the immunodiagnosis of brucellosis in livestock and wildlife","volume":"32","author":"McGiven","year":"2013","journal-title":"Rev. Sci. Tech."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"1204","DOI":"10.1128\/JCM.03185-14","article-title":"Improved serodiagnosis of bovine brucellosis by novel synthetic oligosaccharide antigens representing the capping M epitope elements of Brucella O-polysaccharide","volume":"53","author":"McGiven","year":"2015","journal-title":"J. Clin. Microbiol."},{"key":"ref_88","doi-asserted-by":"crossref","unstructured":"Patra, K.P., Saito, M., Atluri, V.L., Rolan, H.G., Young, B., Kerrinnes, T., Smits, H., Ricaldi, J.N., Gotuzzo, E., and Gilman, R.H. (2014). A protein conjugate approach to develop a monoclonal antibody-based antigen detection test for the diagnosis of human brucellosis. PLoS Negl. Trop. Dis., 8.","DOI":"10.1371\/journal.pntd.0002926"},{"key":"ref_89","first-page":"647","article-title":"Epitope mapping of the Brucella melitensis BP26 immunogenic protein: Usefulness for diagnosis of sheep brucellosis","volume":"10","author":"Verger","year":"2003","journal-title":"Clin. Diagn. Lab. Immunol."},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"1760","DOI":"10.1128\/CVI.05159-11","article-title":"Evaluation of recombinant 10 kDa immunodominant region of BP26 protein of Brucella abortus for specific diagnosis of bovine brucellosis","volume":"18","author":"Tiwari","year":"2011","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"e469","DOI":"10.1016\/j.ijid.2009.06.031","article-title":"Clinical manifestations and complications in 1028 cases of brucellosis: A retrospective evaluation and review of the literature","volume":"14","author":"Buzgan","year":"2010","journal-title":"Int. J. Infect. Dis."},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1097\/00005792-199607000-00003","article-title":"Complications associated with Brucella melitensis infection: A study of 530 cases","volume":"75","author":"Colmenero","year":"1996","journal-title":"Medicine"},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1053\/jinf.1999.0586","article-title":"Brucella bacteraemia: Clinical and laboratory observations in 160 patients","volume":"40","author":"Memish","year":"2000","journal-title":"J. Infect."},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"1895","DOI":"10.3201\/eid1312.070527","article-title":"Changing epidemiology of human brucellosis, Germany, 1962\u20132005","volume":"13","author":"Neubauer","year":"2007","journal-title":"Emerg. Infect. Dis."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/10\/12\/1623\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:47:50Z","timestamp":1760168870000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/10\/12\/1623"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,14]]},"references-count":94,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["pathogens10121623"],"URL":"https:\/\/doi.org\/10.3390\/pathogens10121623","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,12,14]]}}}