Detection of Brucella abortus by immunofluorescence assay using anti outer membrane protein of 19 kDa antibody

Mohammadi, Elham; Golchin, Mehdi

doi:10.17219/acem/85081

Cited by 6 publications

(4 citation statements)

References 35 publications

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“…The FITC-conjugated anti-rOmp19 antibody accurately recognized B. abortus cells and did not bind to E. coli , Salmonella , and Klebsiella . However, the test failed to recognize all Brucella isolates, and it showed positive results when testing half of the isolates of related bacteria used, which is likely due to the presence of common epitopes with heterogeneous microorganisms [ 42 ].…”

Section: Serological Potential Of Brucella Spp Rec...mentioning

confidence: 99%

Brucellosis detection and the role of Brucella spp. cell wall proteins

Bulashev

Eskendirova

2023

Vet World

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Brucellosis remains an endemic zoonotic disease in many developing countries, causing great harm to public health and devastating losses to livestock. One of the main reasons for the low effectiveness of anti-brucellosis measures is the lack of reliable methods for diagnosing infected animals throughout their lifespan. Classical serological tests, such as the tube agglutination test, rose Bengal plate test, and complement fixation test, as well as commercial enzyme-linked immunosorbent assay kits, are based on the detection of antibodies to the cell wall polysaccharide antigens of Brucella spp. smooth strains. As a result, they do not exclude cross-reactions with related bacteria and fail to differentiate between infected and vaccinated animals. Over the past decades, many attempts have been made to identify immunoreactive and pathogen-specific protein antigens. To date, several studies have investigated Brucella spp. recombinant proteins, including cell wall proteins, as the best antigens for diagnosing brucellosis in animals and humans. However, the available results on the specificity and sensitivity of serological tests based on cell wall proteins are ambiguous and sometimes contradictory. This review aims to provide an overview of the current state of knowledge of the diagnostic value of outer membrane and/or periplasmic proteins of Brucella spp. The goal is to identify future developments that may lead to reliable antigens for serological tests. Keywords: Brucella, diagnostics, enzyme-linked immunosorbent assay, outer membrane protein, periplasmic protein.

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Section: Serological Potential Of Brucella Spp Rec...mentioning

confidence: 99%

Brucellosis detection and the role of Brucella spp. cell wall proteins

Bulashev

Eskendirova

2023

Vet World

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“…Blood samples from brucellosis patients were collected at the General Hospital of Agricultural Reclamation Bureau, Harbin, Heilongjiang, China. All blood donor or patient samples were confirmed for negativity or positivity to Brucella infection by Standard Agglutination Test (SAT) and Polymerase Chain Reaction (PCR) (Mohammadi and Golchin, 2018). The PBMCs were prepared according to manufacturer's instructions (Ficoll Pague PLUS, GE Healthcare Life Sciences, USA).…”

Section: Blood Samples From Patients and Blood Donorsmentioning

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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“…Direct detection of Brucella antigens has been hampered by the lack of sensitivity and/or specificity [ 19 ]. These limitations were manifested during either the detection of the whole-cell [ 20 , 21 ] or its subunits [ 22 ]. Nowadays, rapid diagnostics gain researchers’ attention, especially those involving aptamers as a detection agent [ 23 ].…”

Section: Introductionmentioning

confidence: 99%

Generation and Selection of Specific Aptamers Targeting Brucella Species through an Enhanced Cell-SELEX Methodology

El-Husseini

Sayour

Melzer

et al. 2022

IJMS

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Brucellae are Gram-negative, aerobic, non-motile coccobacilli causing brucellosis in man and animals. The disease is one of the most significant yet neglected global zoonoses. Especially in developing countries, brucellosis is causing public health problems and economic losses to private animal owners and national revenues. Composed of oligonucleotides, aptamers are chemical analogues of antibodies that are promising components for developing aptamer-based rapid, sensitive, and specific tests to identify the Brucella group of bacteria. For this purpose, aptamers were generated and selected by an enhanced protocol of cell systematic evolution of ligands by exponential enrichment (cell-SELEX). This enhanced cell-SELEX procedure involved the combination of both conventional and toggle cell-SELEX to boost the specificity and binding affinity to whole Brucella cells. This procedure, combined with high-throughput sequencing of the resulting aptamer pools, comprehensive bioinformatics analysis, and wet lab validation assays, led to the selection of a highly sensitive and specific aptamer for those Brucella species known to circulate in Egypt. The isolated candidate aptamer showed dissociation constant (KD) values of 43.5 ± 11, 61.5 ± 8, and 56 ± 10.8 nM for B. melitensis, B. abortus, and B. suis, respectively. This is the first development of a Brucella-specific aptamer using an enhanced combination of conventional and toggle cell-SELEX to the authors’ best knowledge.

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Detection of Brucella abortus by immunofluorescence assay using anti outer membrane protein of 19 kDa antibody

Abstract: This high-titer antibody could potentially be valuable for the specific diagnostic test of brucellosis.

Cited by 6 publications

References 35 publications

Brucellosis detection and the role of Brucella spp. cell wall proteins

Brucellosis detection and the role of Brucella spp. cell wall proteins

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

Generation and Selection of Specific Aptamers Targeting Brucella Species through an Enhanced Cell-SELEX Methodology

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