Optimization and Validation of Indirect ELISA Using Truncated TssB Protein for the Serodiagnosis of Glanders amongst Equines
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 a...
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Wiley
2014-01-01
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| Series: | The Scientific World Journal |
| Online Access: | http://dx.doi.org/10.1155/2014/469407 |
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| author | Harisankar Singha Praveen Malik Sachin K. Goyal Sandip K. Khurana Chiranjay Mukhopadhyay Vandana K. Eshwara Raj K. Singh |
| author_facet | Harisankar Singha Praveen Malik Sachin K. Goyal Sandip K. Khurana Chiranjay Mukhopadhyay Vandana K. Eshwara Raj K. Singh |
| author_sort | Harisankar Singha |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-c9fa5c84a608485fa474bef55b5d73d3 |
| institution | Kabale University |
| issn | 2356-6140 1537-744X |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | The Scientific World Journal |
| spelling | doaj-art-c9fa5c84a608485fa474bef55b5d73d32025-02-03T01:20:43ZengWileyThe Scientific World Journal2356-61401537-744X2014-01-01201410.1155/2014/469407469407Optimization and Validation of Indirect ELISA Using Truncated TssB Protein for the Serodiagnosis of Glanders amongst EquinesHarisankar Singha0Praveen Malik1Sachin K. Goyal2Sandip K. Khurana3Chiranjay Mukhopadhyay4Vandana K. Eshwara5Raj K. Singh6National Research Centre on Equines, Sirsa Road, Hisar, Haryana 125 001, IndiaNational Research Centre on Equines, Sirsa Road, Hisar, Haryana 125 001, IndiaNational Research Centre on Equines, Sirsa Road, Hisar, Haryana 125 001, IndiaNational Research Centre on Equines, Sirsa Road, Hisar, Haryana 125 001, IndiaDepartment of Microbiology, Kasturba Medical College, Manipal University, Manipal, Karnataka 576104, IndiaDepartment of Microbiology, Kasturba Medical College, Manipal University, Manipal, Karnataka 576104, IndiaNational Research Centre on Equines, Sirsa Road, Hisar, Haryana 125 001, IndiaObjective. 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.http://dx.doi.org/10.1155/2014/469407 |
| spellingShingle | Harisankar Singha Praveen Malik Sachin K. Goyal Sandip K. Khurana Chiranjay Mukhopadhyay Vandana K. Eshwara Raj K. Singh Optimization and Validation of Indirect ELISA Using Truncated TssB Protein for the Serodiagnosis of Glanders amongst Equines The Scientific World Journal |
| title | Optimization and Validation of Indirect ELISA Using Truncated TssB Protein for the Serodiagnosis of Glanders amongst Equines |
| title_full | Optimization and Validation of Indirect ELISA Using Truncated TssB Protein for the Serodiagnosis of Glanders amongst Equines |
| title_fullStr | Optimization and Validation of Indirect ELISA Using Truncated TssB Protein for the Serodiagnosis of Glanders amongst Equines |
| title_full_unstemmed | Optimization and Validation of Indirect ELISA Using Truncated TssB Protein for the Serodiagnosis of Glanders amongst Equines |
| title_short | Optimization and Validation of Indirect ELISA Using Truncated TssB Protein for the Serodiagnosis of Glanders amongst Equines |
| title_sort | optimization and validation of indirect elisa using truncated tssb protein for the serodiagnosis of glanders amongst equines |
| url | http://dx.doi.org/10.1155/2014/469407 |
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