Optimization and validation of indirect ELISA using truncated TssB protein for the serodiagnosis of glanders amongst equines

Harisankar Singha, Praveen Malik, Sachin K. Goyal, Sandip K. Khurana, Chiranjay Mukhopadhyay, Vandana K. Eshwara, Raj K. Singh

Research output: Contribution to journalArticle

9 Citations (Scopus)

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 (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.

Original languageEnglish
Article number469407
JournalThe Scientific World Journal
Volume2014
DOIs
Publication statusPublished - 06-03-2014

Fingerprint

Glanders
Immunosorbents
Serologic Tests
Assays
Burkholderia mallei
Enzyme-Linked Immunosorbent Assay
assay
enzyme
protein
Enzymes
serum
Recombinant Proteins
Proteins
Serum
Horse Diseases
Melioidosis
Disease control
disease control
Amino Acid Sequence
antigen

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Environmental Science(all)

Cite this

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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. 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 (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.",
author = "Harisankar Singha and Praveen Malik and Goyal, {Sachin K.} and Khurana, {Sandip K.} and Chiranjay Mukhopadhyay and Eshwara, {Vandana K.} and Singh, {Raj K.}",
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Optimization and validation of indirect ELISA using truncated TssB protein for the serodiagnosis of glanders amongst equines. / Singha, Harisankar; Malik, Praveen; Goyal, Sachin K.; Khurana, Sandip K.; Mukhopadhyay, Chiranjay; Eshwara, Vandana K.; Singh, Raj K.

In: The Scientific World Journal, Vol. 2014, 469407, 06.03.2014.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Optimization and validation of indirect ELISA using truncated TssB protein for the serodiagnosis of glanders amongst equines

AU - Singha, Harisankar

AU - Malik, Praveen

AU - Goyal, Sachin K.

AU - Khurana, Sandip K.

AU - Mukhopadhyay, Chiranjay

AU - Eshwara, Vandana K.

AU - Singh, Raj K.

PY - 2014/3/6

Y1 - 2014/3/6

N2 - 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 (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.

AB - 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 (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.

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