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Rational design of diagnostic and vaccination strategies for tuberculosis.

Borsuk, S, Seixas, FK, Ramos, DF, Mendum, T, McFadden, J and Dellagostin, O (2012) Rational design of diagnostic and vaccination strategies for tuberculosis. Braz J Infect Dis, 16 (1). pp. 68-73.

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Abstract

The development of diagnostic tests which can readily differentiate between vaccinated and tuberculosis-infected individuals is crucial for the wider utilization of bacillus Calmette-Guérin (BCG) as vaccine in humans and animals. BCG_0092 is an antigen that elicits specific delayed type hypersensitivity reactions similar in size and morphological aspects to that elicited by purified protein derivative, in both animals and humans infected with the tubercle bacilli. We carried out bioinformatics analyses of the BCG_0092 and designed a diagnostic test by using the predicted MHC class I epitopes. In addition, we performed a knockout of this gene by homologous recombination in the BCG vaccine strain to allow differentiation of vaccinated from infected individuals. For that, the flanking sequences of the target gene (BCG_0092)were cloned into a suicide vector. Spontaneous double crossovers, which result in wild type revertants or knockouts were selected using SacB. BCG_0092 is present only in members of the Mycobacterium tuberculosis complex. Eight predicted MHC class I epitopes with potential for immunological diagnosis were defined, allowing the design of a specific diagnostic test. The strategy used to delete the (BCG_0092) gene from BCG was successful. The knockout genotype was confirmed by PCR and by Southern blot. The mutant BCG strain has the potential of inducing protection against tuberculosis without interfering with the diagnostic test based on the use of selected epitopes from BCG_0092.

Item Type: Article
Authors :
NameEmailORCID
Borsuk, SUNSPECIFIEDUNSPECIFIED
Seixas, FKUNSPECIFIEDUNSPECIFIED
Ramos, DFUNSPECIFIEDUNSPECIFIED
Mendum, Tt.mendum@surrey.ac.ukUNSPECIFIED
McFadden, Jj.mcfadden@surrey.ac.ukUNSPECIFIED
Dellagostin, OUNSPECIFIEDUNSPECIFIED
Date : January 2012
Identification Number : https://doi.org/10.1016/S1413-8670(12)70277-5
Depositing User : Symplectic Elements
Date Deposited : 17 May 2017 09:42
Last Modified : 17 May 2017 14:44
URI: http://epubs.surrey.ac.uk/id/eprint/824763

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