Major outer membrane proteins of Brucella spp.: past, present and future

Cloeckaert, Axel; Vizcaı́no, Nieves; Paquet, Jean‐Yves; Bowden, Robert A.; Elzer, Philip H.

doi:10.1016/s0378-1135(02)00211-0

Cited by 159 publications

(126 citation statements)

References 82 publications

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“…Different genetic fragments such as omp31, 16srDNA, omp2 and so on are used in different studies, to identify and differentiate Brucella species and biovars, which could not show ability to differentiate Brucella biovars completely (14,20,21). Therefore, in subsequent studies especially in Cloackaert et al study, omp2a and omp2b fragments and several restriction enzymes were used (15).…”

Section: Discussionmentioning

confidence: 99%

“…In this method, by omp25, omp2 and omp31 loci and all Brucella species can be differentiated and their biovars identified (14). Several studies use these genes to differentiate Brucella species and biovars performed around the World (12,(15)(16)(17), but in Iran, no study was done especially on clinical samples by this method, and it is the first time that a study is done on molecular typing of B. melitensis and B. abortus isolated from blood samples.…”

Section: Introductionmentioning

confidence: 99%

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Molecular Typing of Brucella melitensis and B. abortus from Human Blood Samples Using PCR-RFLP Method

Mirnejad

Mohammadi

Majdi

et al. 2013

Jundishapur J Microbiol

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Background:Brucella is an intracellular parasite of the disease brucellosis throughout the world. Although several molecular typing methods are introduced to find DNA polymorphism that is able to identify Brucella species and biovars, but among these methods, detection of polymorphisms by PCR-RFLP has several advantages including the easy implementation, interpretation and the ease of use for large quantities of samples. Objectives: In the current study, the technique was used for molecular typing of Brucella abortus and B. melitensis that was isolated from human blood samples. Materials and Methods: Blood samples of 160 patients were transferred to Kerman clinical centers with chief complain of acute brucellosis and showed high blood serum level (about 1.80). Their DNAs were extracted by Phenol chloroform method, and the PCR was optimized by using the fragments of designed primers for omp2a and omp2b. Therefore PCR products were restricted by restriction endonuclease as PstI and Hinf1. Finally they were electrophoresed for analyzing the digestion results on agarose gels (2%). Results: In 160 blood samples that were studied with PCR technique, 52 cases obtained bands of 1100 bp for omp2a locus and 1200 bp for omp2b locus from within 52 positive samples by PCR-RFLP method 25 cases (48%) were positive out of which 56% were B. melitensis biovar1 and 44% were B. abortus biovars of 3, 5, 6 or 9. Conclusions:The results of this study showed that PCR-RFLP technique was a fast and applicable method especially for separation, detection and differentiation between species of B. melitensis and B.abortus biovars in blood sample. Also the presented data showed that B. melitensis biovar 1 was the prevalence biovar.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Molecular Typing of Brucella melitensis and B. abortus from Human Blood Samples Using PCR-RFLP Method

Mirnejad

Mohammadi

Majdi

et al. 2013

Jundishapur J Microbiol

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“…Group 1 was identified as minor (94 or 88 kDa). Major Brucella Omps have been classified in group 2 (36 to 38 kDa) and group 3 (Omp25 and Omp31) (11). Moreover, Omp28 was identified as another member of group 3 (26).…”

Section: Praca Oryginalnamentioning

confidence: 99%

Using combined recombinant protein in the diagnosis of bovine brucellosis

Bulashev¹,

Jakubowski²,

Mukantayev³

et al. 2018

Medycyna Weterynaryjna

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SummaryThe aim of this research was to obtain a combined recombinant protein consisting of immunodominant regions of Brucella outer membrane proteins (Omps) with the molecular weights of 25 kDa (Omp25) and 31 kDa (Omp31). The search for candidate proteins was carried out using NCBI PubMed and NCBI GenBank databases. The two most immunodominant regions of Omps were selected. The first region was in a position from 48 to 83 amino acids of Omp31, while the second one was in the position from 180 to 224 amino acids of Omp25. This combined sequence was designated as OmpBm-Ba. The pET32 vector was used for cloning and the expression of the combined recombinant protein in E. coli BL21(DE3). The antigenicity of recombinant OmpBm-Ba (rOmpBm-Ba) as compared with rOmp25 and rOmp31 has been tested on the sera of 109 cattle with positive results to brucellosis according to classical serological tests. The rOmpBm-Ba had a higher antigenicity than the single ones, since it confirmed the presence of Brucella-antibody in all serum samples with positive results of i-ELISA/rOmp25 and/or i-ELISA/rOmp31, as well as additionally revealing 7.3% and 31.2% seropositive animals, respectively. A comparative study of the diagnostic value of i-ELISA/rOmpBm-Ba and conventional tests on blood sera of 24 cattle subjected to a bacteriological examination at slaughter showed a higher reliability of enzyme immunoassay. Further research is necessary to get a more objective evaluation of the diagnostic accuracy of Brucella rOmpBm-Ba by challenging laboratory animals.

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“…16 Among outer membrane proteins of Brucella, molecules with molecular weights of 25, 31 and 36 kDa are the main OMPs. 17 Protein Omp31 was first reported in B. melitensis, it is a protein found in Brucella species except B. abortus and leads to a switch in host immune response to the cellular immunity. 31 kDa cell surface protein of Brucella is highly protected in B. melitensis and it is immunogenic.…”

Section: Introductionmentioning

confidence: 99%

“…31 kDa cell surface protein of Brucella is highly protected in B. melitensis and it is immunogenic. 16,17 In this study, recombinant clone of Omp31 in Escherichia coli was used, which was able to yield high level of protein.…”

Section: Introductionmentioning

confidence: 99%

Development of Surface Plasmon Resonance-Based Immunosensor for Detection ofBrucella melitensis

Saberi¹,

Kamali²,

Taheri³

et al. 2016

Journal of the Brazilian Chemical Society

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A label-free optical immunosensor was developed based on surface plasmon resonance (SPR) method for rapid and selective detection of Brucella melitensis. This immunosensor was constructed by immobilizing capture antibody on 11-mercaptoundecanoic acid modified gold disk. This fabricated immunosensor detected B. melitensis at a concentration range from 10 3 to 10 7 cell mL -1 (R 2 = 0.998) and a detection limit of 100 cell mL -1. Additionally, the kinetic equilibrium dissociation constant (K D ) for assessment of the interaction affinity was calculated as 1.1 × 10 -9 mol L -1 that can be considered as high affinity interaction. This SPR immunosensor provided advantages in term of fast response, label free and accurate detection of B. melitensis in analytical systems.

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Major outer membrane proteins of Brucella spp.: past, present and future

Cited by 159 publications

References 82 publications

Molecular Typing of Brucella melitensis and B. abortus from Human Blood Samples Using PCR-RFLP Method

Molecular Typing of Brucella melitensis and B. abortus from Human Blood Samples Using PCR-RFLP Method

Using combined recombinant protein in the diagnosis of bovine brucellosis

Development of Surface Plasmon Resonance-Based Immunosensor for Detection ofBrucella melitensis

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