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Clinical and vaccine immunology : CVI2012; 19(8); 1193-1198; doi: 10.1128/CVI.00137-12

Evaluation of recombinant proteins of Burkholderia mallei for serodiagnosis of glanders.

Abstract: Glanders is a contagious disease caused by the Gram-negative bacillus Burkholderia mallei. The number of equine glanders outbreaks has increased steadily during the last decade. The disease must be reported to the Office International des Epizooties, Paris, France. Glanders serodiagnosis is hampered by the considerable number of false positives and negatives of the internationally prescribed tests. The major problem leading to the low sensitivity and specificity of the complement fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA) has been linked to the test antigens currently used, i.e., crude preparations of whole cells. False-positive results obtained from other diagnostic tests utilizing crude antigens lead to financial losses to animal owners, and false-negative results can turn a risk into a possible threat. In this study, we report on the identification of diagnostic targets using bioinformatics tools for serodiagnosis of glanders. The identified gene sequences were cloned and expressed as recombinant proteins. The purified recombinant proteins of B. mallei were used in an indirect ELISA format for serodiagnosis of glanders. Two recombinant proteins, 0375H and 0375TH, exhibited 100% sensitivity and specificity for glanders diagnosis. The proteins also did not cross-react with sera from patients with the closely related disease melioidosis. The results of this investigation highlight the potential of recombinant 0375H and 0375TH proteins in specific and sensitive diagnosis of glanders.
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Publication Date: 2012-06-13 PubMed ID: 22695165PubMed Central: PMC3416092DOI: 10.1128/CVI.00137-12Google Scholar: Lookup
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Summary

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This research paper evaluates the use of recombinant proteins from a bacteria, Burkholderia mallei, as a new approach for diagnosing glanders, a contagious equine disease. The study found that two specific proteins provided a 100% accurate diagnosis, improving upon existing methods.

Background

  • Glanders is a contagious disease mainly affecting horses, caused by the Burkholderia mallei bacteria. The disease has seen a resurgence in recent years, prompting an urgent need for more effective diagnostic methods.
  • Currently, the internationally recognised tests for the diagnosis of glanders, the complement fixation test (CFT) and the enzyme-linked immunosorbent assay (ELISA), have significant limitations. The antigens used in these tests, derived from crude preparations of whole bacterial cells, lead to a considerable number of false positive and negative results. This has financial implications for animal owners and could pose a risk to equine health.

Identification of Diagnostic Targets

  • In this study, the researchers used bioinformatics tools to identify potential diagnostic targets for glanders. They focused on specific gene sequences in the B. mallei bacteria.
  • These identified gene sequences were then cloned and expressed as recombinant proteins for further study.

Testing Recombinant Proteins for Glanders Diagnosis

  • The team used the purified recombinant proteins in an indirect ELISA format to test for glanders. This means the proteins were used as ‘reagents’ in the test to see if they could reliably identify a glanders infection.
  • The results revealed that two of the recombinant proteins, called 0375H and 0375TH, had a 100% accuracy in diagnosing glanders. This indicates a significant improvement over the current methods of diagnosis.
  • Furthermore, these proteins did not cross-react with sera from patients with a closely related disease, melioidosis. This suggests that the proteins are specific to glanders and will not give false positive results for other similar diseases.

Conclusion

  • This study suggests that the recombinant proteins 0375H and 0375TH could offer a more specific and sensitive method for diagnosing glanders.
  • These results could have a significant impact on the management of glanders, contributing to a reduction in false diagnoses and helping to more effectively control the disease.

Cite This Article

APA
Pal V, Kumar S, Malik P, Rai GP. (2012). Evaluation of recombinant proteins of Burkholderia mallei for serodiagnosis of glanders. Clin Vaccine Immunol, 19(8), 1193-1198. https://doi.org/10.1128/CVI.00137-12

Publication

Clinical and vaccine immunology : CVI
ISSN: 1556-679X
NlmUniqueID: 101252125
Country: United States
Language: English
Volume: 19
Issue: 8
Pages: 1193-1198

Researcher Affiliations

Pal, Vijai
  • Division of Microbiology, Defence Research and Development Establishment, Gwalior, Madhya Pradesh, India. vjsoni@rediffmail.com
Kumar, Subodh
    Malik, Praveen
      Rai, Ganga Prasad

        MeSH Terms

        • Animals
        • Antigens, Bacterial / genetics
        • Burkholderia mallei / genetics
        • Burkholderia mallei / immunology
        • Computational Biology / methods
        • Enzyme-Linked Immunosorbent Assay / methods
        • Glanders / diagnosis
        • Horses
        • Recombinant Proteins / genetics
        • Sensitivity and Specificity
        • Serologic Tests / methods
        • Veterinary Medicine / methods

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