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The Scientific World Journal
Volume 2014, Article ID 469407, 6 pages
http://dx.doi.org/10.1155/2014/469407
Research Article

Optimization and Validation of Indirect ELISA Using Truncated TssB Protein for the Serodiagnosis of Glanders amongst Equines

1National Research Centre on Equines, Sirsa Road, Hisar, Haryana 125 001, India
2Veterinary Type Culture Collection, National Research Centre on Equines, Sirsa Road, Hisar, Haryana 125 001, India
3Department of Microbiology, Kasturba Medical College, Manipal University, Manipal, Karnataka 576104, India

Received 14 August 2013; Accepted 18 December 2013; Published 3 February 2014

Academic Editors: A. C. Manna and G. A. Rocha

Copyright © 2014 Harisankar Singha et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Objective. To express truncated TssB protein of Burkholderia mallei and to evaluate its diagnostic efficacy for serological detection of glanders among equines. Materials and Methods. In an attempt to develop recombinant protein based enzyme-linked immunosorbent assay (ELISA), N-terminal 200 amino acid sequences of B. mallei TssB protein—a type 6 secretory effector protein—were expressed in prokaryotic expression system. Diagnostic potential of recombinant TssB protein was evaluated in indirect ELISA using a panel of glanders positive (), negative (), and field serum samples (). Cross-reactivity of the assay was assessed with equine disease control serum and human melioidosis positive serum. Results. In comparison to CFT, diagnostic sensitivity and specificity of ELISA were 99.7% and 100%, respectively. Conclusions. The indirect ELISA method using the truncated TssB offered safer and more rapid and efficient means of serodiagnosis of glanders in equines. These data highlight the use of TssB as potential diagnostic antigen for serological diagnosis of glanders.