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Evaluation of recombinant porin (rOmp2a) protein as a potential antigen candidate for serodiagnosis of Human Brucellosis.
- Source :
-
BMC infectious diseases [BMC Infect Dis] 2017 Jul 11; Vol. 17 (1), pp. 485. Date of Electronic Publication: 2017 Jul 11. - Publication Year :
- 2017
-
Abstract
- Background: Brucellosis is an important zoonotic disease caused by different Brucella species and human brucellosis is commonly prevalent in different states of India. Among various Brucella species, B. melitensis is most pathogenic to human and included as category B biothreat which can cause infection through aerosol, cut, wounds in skin and contact with infected animals. The diagnosis of human brucellosis is very important for proper treatment and management of disease as there is no vaccine available for human use. The present study was designed to clone, express and purify immunodominant recombinant omp2a (rOmp2a) porin protein of B. melitensis and to evaluate this new antigen candidate for specific serodiagnosis of human brucellosis by highly sensitive iELISA (indirect enzyme linked immunosorbent assay).<br />Method: Omp2a gene of B. melitensis 16 M strain was cloned and expressed in pET-SUMO expression system. The recombinant protein was purified under denaturing conditions using 8 M urea. The purified recombinant protein was confirmed by western blotting by reacting with anti-HIS antibody. The sero-reactivity of the recombinant protein was also checked by reacting with antisera of experimentally infected mice with B. melitensis 16 M at different time points. Serodiagnostic potential of recombinant porin antigen was tested against 185 clinical serum samples collected from regions endemic to brucellosis in southern part of India by iELISA. The samples were grouped into five groups. Group 1 contained cultured confirmed positive serum samples of brucellosis (n = 15), group 2 contained sera samples from positive cases of brucellosis previously tested by conventional methods of RBPT (n = 28) and STAT (n = 26), group 3 contained sera samples negative by RBPT(n = 36) and STAT (n = 32), group 4 contained sera samples of other febrile illness and PUO case (n = 35) and group 5 contained confirmed negative sera samples from healthy donors (n = 23).<br />Result: The rOmp2a was found to be immunoreactive by iELISA and western blotting. The test showed a sensitivity of 93.75% and specificity of 95.83% when tested against 185 serum samples. For determination of statistical significance between experimental groups and control groups, Student's t test was performed on the data.<br />Conclusion: Omp2a emerges as a potential antigen candidate for serodiagnosis of human brucellosis.
- Subjects :
- Animals
Antibodies, Bacterial blood
Antigens, Bacterial genetics
Antigens, Bacterial immunology
Bacterial Proteins genetics
Bacterial Proteins isolation & purification
Brucella immunology
Brucella pathogenicity
Enzyme-Linked Immunosorbent Assay methods
Humans
India
Mice
Porins genetics
Porins isolation & purification
Recombinant Proteins genetics
Recombinant Proteins immunology
Recombinant Proteins isolation & purification
Sensitivity and Specificity
Bacterial Proteins immunology
Brucellosis diagnosis
Porins immunology
Serologic Tests methods
Subjects
Details
- Language :
- English
- ISSN :
- 1471-2334
- Volume :
- 17
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- BMC infectious diseases
- Publication Type :
- Academic Journal
- Accession number :
- 28693438
- Full Text :
- https://doi.org/10.1186/s12879-017-2588-1