title: Optimization and validation of indirect ELISA using truncated TssB protein for the serodiagnosis of glanders amongst equines.
abstract: OBJECTIVE
  To express truncated TssB protein of Burkholderia mallei and to evaluate its diagnostic efficacy for serological detection of glanders among equines.
  
  
  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 (n = 49), negative (n = 30), and field serum samples (n = 1811). 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.
authors.affiliation: National Research Centre on Equines, Sirsa Road, Hisar, Haryana 125 001, India.
authors.forename: Harisankar
authors.identifier: 0000-0003-4607-9212
authors.initials: H
authors.lastname: Singha
authors.affiliation: National Research Centre on Equines, Sirsa Road, Hisar, Haryana 125 001, India ; Veterinary Type Culture Collection, National Research Centre on Equines, Sirsa Road, Hisar, Haryana 125 001, India.
authors.forename: Praveen
authors.identifier: 0000-0001-7075-8106
authors.initials: P
authors.lastname: Malik
authors.affiliation: National Research Centre on Equines, Sirsa Road, Hisar, Haryana 125 001, India.
authors.forename: Sachin K
authors.initials: SK
authors.lastname: Goyal
authors.affiliation: National Research Centre on Equines, Sirsa Road, Hisar, Haryana 125 001, India.
authors.forename: Sandip K
authors.initials: SK
authors.lastname: Khurana
authors.affiliation: Department of Microbiology, Kasturba Medical College, Manipal University, Manipal, Karnataka 576104, India.
authors.forename: Chiranjay
authors.initials: C
authors.lastname: Mukhopadhyay
authors.affiliation: Department of Microbiology, Kasturba Medical College, Manipal University, Manipal, Karnataka 576104, India.
authors.forename: Vandana K
authors.initials: VK
authors.lastname: Eshwara
authors.affiliation: National Research Centre on Equines, Sirsa Road, Hisar, Haryana 125 001, India.
authors.forename: Raj K
authors.initials: RK
authors.lastname: Singh
mesh_terms: D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D042726:Burkholderia mallei / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005896:Glanders / Q000175:diagnosis*; D006734:Horse Diseases / Q000175:diagnosis*; D006736:Horses; D008969:Molecular Sequence Data; D012689:Sequence Homology, Nucleic Acid; D012698:Serologic Tests