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      The Rose Bengal Test in Human Brucellosis: A Neglected Test for the Diagnosis of a Neglected Disease

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          Abstract

          Brucellosis is a highly contagious zoonosis affecting livestock and human beings. The human disease lacks pathognomonic symptoms and laboratory tests are essential for its diagnosis. However, most tests are difficult to implement in the areas and countries were brucellosis is endemic. Here, we compared the simple and cheap Rose Bengal Test (RBT) with serum agglutination, Coombs, competitive ELISA, Brucellacapt, lateral flow immunochromatography for IgM and IgG detection and immunoprecipitation with Brucella proteins. We tested 208 sera from patients with brucellosis proved by bacteriological isolation, 20 contacts with no brucellosis, and 1559 sera of persons with no recent contact or brucellosis symptoms. RBT was highly sensitive in acute and long evolution brucellosis cases and this related to its ability to detect IgM, IgG and IgA, to the absence of prozones, and to the agglutinating activity of blocking IgA at the pH of the test. RBT was also highly specific in the sera of persons with no contact with Brucella. No test in this study outperformed RBT, and none was fully satisfactory in distinguishing contacts from infected patients. When modified to test serum dilutions, a diagnostic titer >4 in RBT resulted in 87.4% sensitivity (infected patients) and 100% specificity (contacts). We discuss the limitations of serological tests in the diagnosis of human brucellosis, particularly in the more chronic forms, and conclude that simplicity and affordability of RBT make it close to the ideal test for small and understaffed hospitals and laboratories.

          Author Summary

          The Rose Bengal Test (RBT) for brucellosis serological diagnosis was adapted to test serum dilutions and its usefulness evaluated using sera of Brucella culture positive patients, persons with contact with Brucella but no symptoms, veterinarians accidentally injected with vaccine Rev 1 who had not developed the disease and normal persons. Using the standard protocol, RBT was not outperformed by more sophisticated and expensive tests (serum agglutination, Coombs, competitive ELISA, Brucellacapt, and lateral flow immunochromatography for IgM and IgG detection) in identifying Brucella infected patients. All tests failed to discriminate with total specificity the sera from contacts or Rev 1 injected individuals. However, none of these sera was positive in the modified RBT adapted to test serum dilutions at titers higher than 1>4. When there is suspicion of brucellosis, RBT is recommended as the first test and, depending upon the titer, a positive result does not need confirmation by other (usually more expensive, sophisticated and time consuming) tests.

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          Most cited references58

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          Human brucellosis.

          Human brucellosis still presents scientists and clinicians with several challenges, such as the understanding of pathogenic mechanisms of Brucella spp, the identification of markers for disease severity, progression, and treatment response, and the development of improved treatment regimens. Molecular studies have shed new light on the pathogenesis of Brucella spp, and new technologies have permitted the development of diagnostic tools that will be useful in developing countries, where brucellosis is still a very common but often neglected disease. However, further studies are needed to establish optimum treatment regimens and local and international control programmes. This Review summarises current knowledge of the pathogenic mechanisms, new diagnostic advances, therapeutic options, and the situation of developing countries in regard to human brucellosis.
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            Brucellosis: an overview.

            M Corbel (1997)
            Brucellosis remains a major zoonosis worldwide. Although many countries have eradicated Brucella abortus from cattle, in some areas Brucella melitensis has emerged as a cause of infection in this species as well as in sheep and goats. Despite vaccination campaigns with the Rev 1 strain, B. melitensis remains the principal cause of human brucellosis. Brucella suis is also emerging as an agent of infection in cattle, thus extending its opportunities to infect humans. The recent isolation of distinctive strains of Brucella from marine mammals has extended its ecologic range. Molecular genetic studies have demonstrated phylogenetic affiliation to Agrobacterium, Phyllobacterium, Ochrobactrum, and Rhizobium. Polymerase chain reaction and gene probe development may provide more effective typing methods. Pathogenicity is related to production of lipopolysaccharides containing a poly N-formyl perosamine O chain, CuZn superoxide dismutase, erythrlose phosphate dehydrogenase, stress-induced proteins related to intracellular survival, and adenine and guanine monophosphate inhibitors of phagocyte functions. Protective immunity is conferred by antibody to lipopolysaccharide and T-cell-mediated macrophage activation triggered by protein antigens. Diagnosis still centers on isolation of the organism and serologic test results, especially enzyme immunoassay, which is replacing other methods. Polymerase chain reaction is also under evaluation. Therapy is based on tetracyclines with or without rifampicin, aminoglycosides, or quinolones. No satisfactory vaccines against human brucellosis are available, although attenuated purE mutants appear promising.
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              Brucellosis in Humans and Animals

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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS Negl Trop Dis
                plos
                plosntds
                PLoS Neglected Tropical Diseases
                Public Library of Science (San Francisco, USA )
                1935-2727
                1935-2735
                April 2011
                19 April 2011
                : 5
                : 4
                : e950
                Affiliations
                [1 ]Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad de Navarra, Pamplona, Spain
                [2 ]Departamento de Microbiología, Hospital Universitari de Bellvitge, Universidad de Barcelona (IDIBELL), Barcelona, Spain
                [3 ]Departamento de Enfermedades Infecciosas, Hospital Universitari de Bellvitge, Universidad de Barcelona (IDIBELL), Barcelona, Spain
                University of California San Diego School of Medicine, United States of America
                Author notes

                Conceived and designed the experiments: RD JA. Performed the experiments: RD AC. Analyzed the data: RD AC JA IM. Contributed reagents/materials/analysis tools: AC. Wrote the paper: IM.

                Article
                10-PNTD-RA-1202R3
                10.1371/journal.pntd.0000950
                3079581
                21526218
                e966a47b-e503-49aa-973b-8280574b9c18
                Díaz et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
                History
                : 25 May 2010
                : 23 December 2010
                Page count
                Pages: 7
                Categories
                Research Article
                Infectious Diseases/Bacterial Infections
                Microbiology/Medical Microbiology

                Infectious disease & Microbiology
                Infectious disease & Microbiology

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