A systematic review of current immunological tests for the diagnosis of cattle brucellosis

Ducrotoy, Marie J.; Muñoz, Pilar M.; Conde-Álvarez, Raquel; Blasco, J.M.; Moriyón, Ignacio

doi:10.1016/j.prevetmed.2018.01.005

Cited by 76 publications

(113 citation statements)

References 109 publications

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“…Meanwhile, the iELISA is considered to be sensitive and could be used as a single diagnostic criterion at standardized labs [30]. However, RBPT remains an adequate screening test based upon the disease epidemiology, purpose of the diagnostic criteria and availability of the resources [31,32]. Therefore, we tested our sera by both tests and determined the possible agreement between these two tests (Tables 5 and 6).…”

Section: Discussionmentioning

confidence: 99%

Serological and Molecular Detection of Bovine Brucellosis at Institutional Livestock Farms in Punjab, Pakistan

Jamil

Melzer

Saqib

et al. 2020

IJERPH

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Bovine brucellosis remains a persistent infection in ruminants in Pakistan. A total of 828 (409 buffaloes and 419 cattle) sera were collected from 11 institutional-owned livestock farms in Punjab, Pakistan. The samples were tested by rose bengal plate agglutination test (RBPT) and indirect enzyme-linked immunosorbent assay (iELISA). The seroprevalence along with 95% confidence interval (CI) was determined. Univariable and multivariable analysis of the epidemiological background data was conducted and odds ratio (OR) was calculated to understand any association between the risk factors and the seroprevalence. An overall seroprevalence of 3.9% (Positive/Tested = 32/828) and 3.3% (27/828) was detected by RBPT and iELISA, respectively. The seroprevalence of 5.6% (CI 3.6–8.3) and 4.7%, (CI 2.8–7.2) and the odds ratio of 2.63 (CI 1.20–5.77) and 2.50 (CI 1.08–5.78) for testing positive by RBPT and iELISA, respectively were significantly higher (p < 0.05) in buffaloes than in cattle. Breed, sex, history of abortion and retention of fetal membranes (RFM) in the animals were not found statistically significantly associated with the infection. RBPT and iELISA based results agreed almost perfect (k = 0.877). In total, Brucella abortus-DNA (9/27) was amplified from seropositive samples by real-time polymerase chain reaction. This study identified for the first time the etiological agents of brucellosis at a molecular level at institutional-owned livestock farms in Pakistan.

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Section: Discussionmentioning

confidence: 99%

Serological and Molecular Detection of Bovine Brucellosis at Institutional Livestock Farms in Punjab, Pakistan

Jamil

Melzer

Saqib

et al. 2020

IJERPH

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“…By aligning Brucella Omp25 aa sequence with Ochrobactrum anthropi and Rhizobiales 63-22 strains, the high homologous sequences are observed including the linear epitopes (P2, P4, and P7) within Brucella Omp25 recognized by mAbs 4A12/4F10, 8F3, and 2B10 (Figure S3), which indicate the cross-reactivity of these mAbs may exist in other Alphaproteobacteria. B. melitensis, B. abortus, and B. suis are three major pathogenic strains for human brucellosis transmitted from infected sheep and goats, cattle, or pigs Ducrotoy et al, 2018). Among five mAbs, 8F3, 4A12, and 6C12 had higher relative affinity (>1) reacted with Omp25 of five strains (M5-90, S19, 104M, 2308, and S2) from three Brucella in various immunoassays.…”

Section: Discussionmentioning

confidence: 99%

“…Brucellosis is a severe zoonotic disease distributed worldwide, especially in the developing world . Sheep, goats, cattle, and pigs infected by Brucella species are the main sources of human brucellosis (Ducrotoy et al, 2018). In the past 10 years, human brucellosis has increased rapidly in China.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Reactivity of Monoclonal Antibodies Against Omp25 of Brucella spp.

Yang

et al. 2020

Front. Cell. Infect. Microbiol.

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Brucellosis is a serious zoonosis occurring mainly in developing countries, and its diagnosis is largely dependent on serologic detection and bacterial culture. In this study, we developed the murine monoclonal antibodies (mAbs) against a conserved and major outer membrane protein 25 (Omp25) of Brucella species (B. spp.) for use in clinical diagnosis. The mAbs to Omp25 were produced by hybridoma technique, which were utilized for developing various immunoassays for detection of Brucellae, including Western blot (WB), enzyme-linked immunosorbent assay (ELISA), immunochemical staining (ICS), immunofluorescence staining (IFS), and flow cytometry assay (FCM). A number of five mAbs (2B10, 4A12, 4F10, 6C12, and 8F3) specific to Omp25 were selected, including 2 IgG1, 2 IgG2a, and 1 IgG2b. Among them, mAbs 6C12, 8F3, and 4A12 reacted highly with B. melitensis (M5-90), B. abortus (S19, 104M, and 2308), and B. suis strain (S2). No cross-reactivity with Yersinia enterocolitica O:9, Salmonella spp., and Escherichia coli was found. By mapping Omp25 epitopes, mAb 6C12 was found as reacting with a semi-conformational epitope, and mAbs 4A12 and 8F3 as recognizing a different linear epitope, respectively. The paired mAbs were tested for detecting Brucella species, suggesting that 8F3 was suitable for solid phase capture and 6C12 or 4A12 was suitable for conjugation with HRP for detection of Brucella Omp25 in ELISA. The FCM was established by mAb 6C12 for detecting intracellular Brucellae-infected peripheral blood mononuclear cells (PBMCs) from brucellosis patients. In conclusion, mAbs against Omp25 are precious reagents for detection of Brucellae in clinical samples with various immunoassays. mAb 6C12-based FCM could be potentially used for the monitoring of therapeutic efficacy for brucellosis in clinical practice.

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“…Reducing the global burden of animal brucellosis will decrease the incidence of the disease in humans and compliance to control programs together with accurate diagnosis are instrumental to achieve this goal. The use of different strategies to control the disease, as well as the lack of diagnostic tests able to unequivocally diagnose the infection, impairs the effectiveness of control programs [1,8]. In addition, the presence of FPSRs imposes the use of combined seroepidemiologic methods and the development of better tests to implement diagnosis in animals.…”

Section: Discussionmentioning

confidence: 99%

“…Bacterial isolation and identification is clearly the gold standard diagnostic method but it is time consuming and impractical since it is performed on organs from slaughtered animals; in addition, the low isolation rate from infected tissues often results in false negatives [1,5]. Serologic assays are rapid and simple systems to detect infected animals; several tests based on different principles have been developed worldwide to reach a good level of specificity and sensitivity although neither ideal nor unique serological test is available to precisely diagnose animal brucellosis [6][7][8]. Mediterranean countries are not brucellosis-free; in particular, in Italy the disease occurs in ruminants with low prevalence in Southern regions and, since vaccination is not allowed, the test-and-slaughter strategy together with sero-epidemiological surveillance programs are in force to control the disease in these areas.…”

Section: Introductionmentioning

confidence: 99%

Brucella melitensis B115-based ELISA to unravel false positive serologic reactions in bovine brucellosis: a field study

et al. 2020

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Background: Brucellosis is a zoonosis whose incidence is not declining worldwide despite the global effort to control the disease. Accurate and precise diagnosis is a crucial step in any prophylaxis program but single tests to unequivocally detect animals infected with Brucella spp. are currently unavailable. In Italy, serological diagnosis of bovine brucellosis is performed with two official tests: a rapid agglutination test (i.e., Rose Bengal Plate test, RBPT) and a complement fixation test (CFT) that detect antibodies directed mainly to the smooth lipopolysaccharide (S-LPS). Neither of the two tests is able to avoid the detection of false positive serological reactions (FPSRs) caused by bacteria sharing S-LPS components with Brucella spp. and responsible for the single reactors (SR) phenomenon. A B. melitensis R strain-based ELISA showed a good diagnostic performance in unravelling FP animals; however, since a limited number of animals were analyzed in that study, a large field study was conducted here to discriminate between Brucella-infected from FP animals, with the final aim of reducing the unnecessary slaughter of the latter. An ELISA based on a R strain of Brucella, i.e., Brucella melitensis B115, was employed to measure specific IgG responses in a collection of bovine sera (n = 648). Sera were obtained from 180 farms (either officially brucellosisfree or not brucellosis-free) recruited during an extended period of time (2014)(2015)(2016)(2017)(2018) and were preliminarily assayed with the official tests by the Italian Reference Centers and then subjected to the ELISA. Results: Negative sera, when subjected to the ELISA, gave O.D. values below the cutoff; SR sera, i.e. RBPT positive and CFT negative, as well as double positive (DP) sera, i.e. RBPT and CFT positive, gave O.D. values that were below the cutoff. All positive sera, i.e. from Brucella-infected animals, were RBPT positive and CFT positive (ICFTU ranging from 20 to 1280) and gave ELISA O.D. values above the cutoff. Conclusions: The B. melitensis B115-based ELISA systematically unravelled all false positive (FP) sera while confirming the diagnosis in Brucella-infected animals. Thus, the test employed in the present study may complement the official assays to avoid the costly slaughter of FP animals.

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A systematic review of current immunological tests for the diagnosis of cattle brucellosis

Cited by 76 publications

References 109 publications

Serological and Molecular Detection of Bovine Brucellosis at Institutional Livestock Farms in Punjab, Pakistan

Serological and Molecular Detection of Bovine Brucellosis at Institutional Livestock Farms in Punjab, Pakistan

Evaluation of Reactivity of Monoclonal Antibodies Against Omp25 of Brucella spp.

Brucella melitensis B115-based ELISA to unravel false positive serologic reactions in bovine brucellosis: a field study

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