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Cloning, expression and purification of virB10 protein of Brucella melitensis and evaluation of its role as a serological marker for Brucella infection in experimental and natural host.
- Source :
-
Protein expression and purification [Protein Expr Purif] 2018 May; Vol. 145, pp. 53-58. Date of Electronic Publication: 2018 Jan 03. - Publication Year :
- 2018
-
Abstract
- Brucellosis is a zoonotic disease caused by various species of the genus Brucella. The control of this disease mainly depends on its accurate and early diagnosis. Culture methods employed for diagnosis are time consuming and require well equipped biosafety level 3 laboratories and hence serological tests are favored alternative for brucellosis diagnosis. At present serological diagnosis is based on LPS (lipopolysaccharide) which is less specific as it shows cross reactivity with other gram-negative bacteria. There is a need to develop serological diagnostic assay based on purified recombinant antigen of Brucella. T4SS (Type IV Secretion System) is an important virulent factor of Brucella and required for infection suggesting their expression in vivo and can be targeted as serological marker for infection. To test this concept, the present study is designed to clone, express and purify virB10 gene of Brucella T4SS under denaturing conditions and to evaluate its use as serological marker of Brucella infection. The immunoreactivity of this recombinant antigen was checked with antisera collected after experimental infection in Balb/C mice with B. melitensis 16M, BR31 (human clinical isolate) and Y. enterocolitica O:9. The recombinant protein was also tested against a panel of 46 bovine sera samples collected from Leh, India. Antibody response against VirB10 was detected in experimental and natural host suggesting that it can be explored as potential target for serodiagnosis of Brucella infection.<br /> (Copyright © 2018 Elsevier Inc. All rights reserved.)
- Subjects :
- Animals
Antigens, Bacterial immunology
Antigens, Bacterial isolation & purification
Brucella melitensis immunology
Brucellosis immunology
Brucellosis metabolism
Cattle
Gene Expression
Humans
Mice
Recombinant Proteins genetics
Recombinant Proteins immunology
Recombinant Proteins isolation & purification
Sensitivity and Specificity
Antibodies, Bacterial blood
Antigens, Bacterial genetics
Brucella melitensis metabolism
Brucellosis diagnosis
Serologic Tests methods
Subjects
Details
- Language :
- English
- ISSN :
- 1096-0279
- Volume :
- 145
- Database :
- MEDLINE
- Journal :
- Protein expression and purification
- Publication Type :
- Academic Journal
- Accession number :
- 29305897
- Full Text :
- https://doi.org/10.1016/j.pep.2017.12.014