Immunogenic proteins of Brucella abortus to minimize cross reactions in brucellosis diagnosis

Ko, Kyung Yuk; Kim, Jongwan; Her, Moon; Kang, Sung-Il; Jung, Suk-Chan; Cho, Dong Hee; Kim, Jiyeon

doi:10.1016/j.vetmic.2011.11.011

Cited by 42 publications

(40 citation statements)

References 24 publications

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“…AtpD immunoreactivity has also been reported in M. mycoides subsp . mycoides SC, Brucella abortus, and Brucella melitensis [24, 37, 38]. Similarly, the antigenic properties of TktA and Ef-Tu have been reported in several mycoplasma species [24, 25, 35, 39].…”

Section: Discussionmentioning

confidence: 86%

Immunoproteomic identification of MbovP579, a promising diagnostic biomarker for serological detection of Mycoplasma bovis infection

et al. 2016

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A lack of knowledge regarding the antigenic properties of Mycoplasma bovis proteins prevents the effective control of bovine infections using immunological approaches. In this study, we detected and characterized a specific and sensitive M. bovis diagnostic biomarker. After M. bovis total proteins and membrane fractions were separated with two dimensional gel electrophoresis, proteins reacting with antiserawere detected using MALDI-TOF MS. Thirty-nine proteins were identified, 32 of which were previously unreported. Among them, immunoinformatics predicted eight antigens, encoded by Mbov_0106, 0116, 0126, 0212, 0275, 0579, 0739, and 0789, to have high immunological value. These genes were expressed in E. coli after mutagenesis of UGA to UGG using overlap extension PCR. A lipoprotein, MbovP579, encoded by a functionally unknown gene, was a sensitive and specific antigen for detection of antibodies in sera from both M. bovis-infected and vaccinated cattle. The specificity of MbovP579 was confirmed by its lack of cross-reactivity with other mycoplasmas, including Mycoplasma agalactiae. An iELISA based on rMbovP579 detected seroconversion 7 days post-infection (dpi). The ELISA had sensitivity of 90.2% (95% CI: 83.7%, 94.3%) and a specificity of 97.8% (95% CI: 88.7%, 99.6%) with clinical samples. Additional comparative studies showed that both diagnostic and analytic sensitivities of the ELISA were higher than those of a commercially available kit (p<0.01). We have thus detected and characterized the novel antigen, MbovP579, and established an rMbovP579-based ELISA as a highly sensitive and specific method for the early diagnosis of M. bovis infection.

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Section: Discussionmentioning

confidence: 86%

Immunoproteomic identification of MbovP579, a promising diagnostic biomarker for serological detection of Mycoplasma bovis infection

et al. 2016

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“…Outer membrane proteins (OMPs) of Brucella spp. have been the focus of vaccine development and the diagnosis of brucellosis (Cloeckaert et al, 2001;Gupta et al, 2010;Ko et al, 2012). OMP28 is considered as one of the outer membrane proteins of Brucella (Cha et al, 2012) and has been identified as an important diagnostic antigen in brucellosis (Seco-Mediavilla et al, 2003;Poester et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Expression, purification and immunochemical characterization of recombinant OMP28 protein of <i>Brucella</i> species

Manat¹,

Shustov²,

Evtehova³

et al. 2016

Open Vet J.

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Brucellosis is the lion's share of infectious disease of animals and it has a particular socio-economic importance for the Republic of Kazakhstan. Sixty percent of epizootic outbreaks of brucellosis identified in the Commonwealth of Independent States (CIS) originated from Kazakhstan in recent years. Definitive diagnosis of brucellosis remains a difficult task. Precisely for this reason, we evaluated a purified recombinant out membrane protein 28 (rOMP28) of Brucella species (Brucella spp.) produced in Escherichia coli (E. coli) as a diagnostic antigen in an Indirect ELISA (I-ELISA) for bovine brucellosis. The gene encoding OMP28 was synthesized using a two-round PCR procedure. In order to produce the rOMP28, the de novo synthesized DNA was cloned into the expression vector pET-22b(+). Then, the rOMP28 was expressed in E. coli system and characterized in the present study. We further estimated the diagnostic potential of purified rOMP28 of Brucella spp. for screening bovine sera. To determine if rOMP28 has a valuable benefit for use in the serodiagnosis of bovine brucellosis, rOMP28-based I-ELISA was performed. Brucella spp. positive (n=62) and Brucella spp. negative (n=28) samples from tube agglutination test (TAT) were positive (n=59) and negative (n=27) by I-ELISA, respectively. These findings show that the rOMP28 of Brucella spp. could be a good candidate for improving serological diagnostic methods for bovine brucellosis.

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“…The renovation of these parallels can notably improve diagnostic tools and vaccine development against Brucella infections. In the last few decades, the low specificity of diagnosis due to cross-reacting bacteria and the poor efficiency of live attenuated vaccine with Brucella strains has impeded the development of a treatment for this painful disease (Ko et al, 2012). In practice, diverse modifications for the diagnosis of brucellosis have been investigated to overcome the limitations arising from the immune response against Brucella infection (McGiven et al, 2008).…”

Section: Discussionmentioning

confidence: 99%

Proteomic analyses of the time course responses of mice infected withBrucella abortus544 reveal immunogenic antigens

Lee

Simborio

Reyes

et al. 2014

FEMS Microbiol Lett

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Brucellosis is a major zoonotic disease caused by pathogens of the genus Brucella. The eradication of brucellosis in domestic animals, associated with the prevention of human infection, can be attained through accurate diagnosis. However, the conventional serological diagnosis of brucellosis has limitations, particularly in detecting the infection period. Accordingly, the aim of this study was to determine reliable immunogenic proteins to detect Brucella abortus infection according to time course responses to aid in the appropriate management of this disease. Proteomic identification through two-dimensional electrophoresis (2DE), followed by immunoblotting, revealed 13, 24, and 55 immunodominant B. abortus 544 proteins that were reactive to sera from experimentally infected mice at early (10 days), middle (30 days), and late (60 days) infection periods, respectively. After excluding several spots reactive to sera from Yersinia enterocolitica O:9-infected and noninfected mice, 17 of the 67 immunodominant proteins were identified through MALDI-TOF MS. Consequently, the identified proteins showed time course-dependent immunogenicity against Brucella infection. Thus, the results of this study suggest that the production of immunogenic proteins during infection periods improves the diagnosis and discovery of vaccine candidates.

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Immunogenic proteins of Brucella abortus to minimize cross reactions in brucellosis diagnosis

Cited by 42 publications

References 24 publications

Immunoproteomic identification of MbovP579, a promising diagnostic biomarker for serological detection of Mycoplasma bovis infection

Immunoproteomic identification of MbovP579, a promising diagnostic biomarker for serological detection of Mycoplasma bovis infection

Expression, purification and immunochemical characterization of recombinant OMP28 protein of <i>Brucella</i> species

Proteomic analyses of the time course responses of mice infected withBrucella abortus544 reveal immunogenic antigens

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