Use of a recombinant burkholderia intracellular motility a protein for immunodiagnosis of glanders.
Abstract: Glanders, caused by the Gram-negative, nonmotile bacterium Burkholderia mallei, is a contagious and highly fatal disease of equines. During the last decade, the number of glanders outbreaks has increased steadily. The disease also has high zoonotic significance and B. mallei is listed biological warfare agent. The complement fixation test (CFT) is a routinely used and internationally recognized test to screen equine sera for the glanders. However, discrepant results have been observed using the CFT. The low sensitivity and specificity of the CFT and enzyme-linked immunosorbent assay (ELISA) have been linked to the use of crude test antigens. We expressed a novel recombinant Burkholderia intracellular motility A (rBimA) protein in Escherichia coli for the diagnosis of equine glanders. Purified rBimA was used in an indirect ELISA format. All of the 21 true-positive serum samples used in the study tested positive, whereas only 17 of the 1,524 potentially negative sera tested positive by indirect ELISA, thus exhibiting 100% sensitivity and 98.88% specificity. Also, rBimA protein did not react with melioidosis patient and normal healthy human serum samples, showing its high specificity. The developed assay can be used as a simple and rapid tool for diagnosis of glanders in equine serum samples. An Indian patent (1328/DEL/2010) has been filed for the reagent.
Publication Date: 2011-07-13 PubMed ID: 21752949PubMed Central: PMC3165212DOI: 10.1128/CVI.05185-11Google Scholar: Lookup
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Summary
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The research paper details the development and validation of a novel diagnostic tool for glanders, a fatal disease caused by the bacterium Burkholderia mallei, affecting equines and also posing a risk to humans. This new tool, based on the recombinant Burkholderia intracellular motility A (rBimA) protein, exhibited a high level of sensitivity and specificity in differentiating between true positive and negative serum samples.
Context
- The paper discusses an infectious disease known as glanders, a highly consequential infection among horses, caused by the bacterium Burkholderia mallei.
- The infection also has significant zoonotic implications, meaning it can be transferred from animals to humans.
- Inaccurate results with the commonly used diagnostic method, the complement fixation test (CFT), drove the researchers to explore a new avenue for diagnosing glanders.
Development of the Diagnostic Tool
- The researchers identified the Burkholderia intracellular motility A (BimA) protein as a potential target for creating a diagnostic test. This protein was artificially recreated (recombinant), a variant named rBimA.
- This rBimA variant was expressed in E. Coli, purified and used in a diagnostic format known as an indirect enzyme-linked immunosorbent assay (ELISA).
Validation of the Diagnostic Tool
- The researchers then tested the performance of the rBimA-based ELISA using 21 true-positive serum samples and 1,524 possibly negative serum samples.
- Results showed that all 21 true-positive samples tested positive, while only 17 out of the 1,524 potentially negative samples tested positive.
- This translates to a sensitivity of 100% (ability to correctly identify positive cases) and a specificity of 98.88% (ability to correctly identify negative cases).
Conclusion and Future Applications
- Because the rBimA protein did not react with serum from patients with melioidosis (a different disease caused by a related bacterium) or with serum from healthy individuals, the researchers believe it shows a high specificity for glanders diagnosis.
- The developed assay can serve as a rapid and effective tool to diagnose glanders within equine serum samples, overcoming limitations of currently used tests.
- The reagent has been filed for patent rights in India (1328/DEL/2010).
Cite This Article
APA
Kumar S, Malik P, Verma SK, Pal V, Gautam V, Mukhopadhyay C, Rai GP.
(2011).
Use of a recombinant burkholderia intracellular motility a protein for immunodiagnosis of glanders.
Clin Vaccine Immunol, 18(9), 1456-1461.
https://doi.org/10.1128/CVI.05185-11 Publication
Researcher Affiliations
- Division of Microbiology, Defence Research and Development Establishment, Jhansi Road, Gwalior-474 002, India. subodh_kumar@email.com
MeSH Terms
- Amino Acid Sequence
- Animals
- Antibodies, Bacterial / blood
- Bacterial Proteins / genetics
- Bacterial Proteins / immunology
- Bacterial Proteins / metabolism
- Burkholderia mallei / genetics
- Burkholderia mallei / immunology
- Escherichia coli / genetics
- Escherichia coli / metabolism
- Glanders / diagnosis
- Glanders / immunology
- Glanders / microbiology
- Horse Diseases / diagnosis
- Horse Diseases / immunology
- Horse Diseases / microbiology
- Horses
- Humans
- Microfilament Proteins / genetics
- Microfilament Proteins / immunology
- Microfilament Proteins / metabolism
- Molecular Sequence Data
- Recombinant Proteins / genetics
- Recombinant Proteins / immunology
- Recombinant Proteins / metabolism
- Sensitivity and Specificity
References
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