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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

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

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

        References

        This article includes 33 references
        1. Al-Ani FK. Glanders in horses: a review.. Sud. J. Vet. Sci. Anim. Husb. 1993 32:1–10.
        2. Al-Ani FK, Al-Rawashdeh OF, Ali AH, Hassan FK. Glanders in horses: clinical, biochemical and serological studies in Iraq.. Vet. Arch. 1998 68:155–162.
        3. Anuntagool N, Sirisinha S. Antigenic relatedness between Burkholderia pseudomallei and Burkholderia mallei.. Microbiol. Immunol. 2002 46:143–150.
          pubmed: 12008922
        4. Arun S. Equine glanders in Turkey.. Vet. Rec. 1999 144:255–258.
          pubmed: 10209817
        5. Bazargani TT, Tadjbakhs H, Badii A, Zahraei T. The outbreak of glanders in some racehorses in three states of Iran.. J. Equine Vet. Sci. 1996 16:232–236.
        6. Boyden SV. Absorption by erythrocytes of antigens of Pf. mallei and Pf. whitmori.. Proc. Soc. Exp. Biol. Med. 1950 73:289.
        7. Brett PJ, DeShazer D, Woods DE. Burkholderia thailandensis sp. nov., a Burkholderia pseudomallei-like species.. Int. J. Syst. Bacteriol. 1998 48:317–320.
          pubmed: 9542103
        8. Cravitz L, Miller WR. Immunologic studies with Malleomyces mallei and Malleomyces pseudomallei I: serological relationships between M. mallei and M. pseudomallei.. J. Infect. Dis. 1950 86:46–51.
          pubmed: 15402290
        9. Elschner MC. Burkholderia mallei infection in a horse imported from Brazil.. Equine Vet. Educ. 2009 21:147–150.
        10. Elschner MC. Use of a Western blot technique for the serodiagnosis of glanders.. BMC Vet. Res. 2011 7:4.
          pmc: PMC3034690pubmed: 21247488
        11. Godoy D. Multilocus sequence typing and evolutionary relationships among the causative agents of melioidosis and glanders, Burkholderia pseudomallei and Burkholderia mallei.. J. Clin. Microbiol. 2003 41:2068–2079.
          pmc: PMC154742pubmed: 12734250
        12. Holden MTG. Genomic plasticity of the causative agent of melioidosis, Burkholderia pseudomallei.. Proc. Natl. Acad. Sci. U. S. A. 2004 101:14240–14245.
          pmc: PMC521101pubmed: 15377794
        13. Jana AM, Gupta AK, Pandya G, Verma RD, Rao KM. Rapid diagnosis of glanders in equines by counter-immuno-electrophoresis.. Indian Vet. J. 1982 59:5–9.
        14. Katz J, Dewald R, Nicholson J. Procedurally similar competitive immunoassay systems for the serodiagnosis of Babesia equi, Babesia caballi, Trypanosoma equiperdum and Burkholderia mallei infection in horses.. J. Vet. Diagn. Invest. 2000 12:46–50.
          pubmed: 10690775
        15. Kumar S. Use of a recombinant Burkholderia intracellular motility A protein for immunodiagnosis of glanders.. Clin. Vaccine Immunol. 2011 18:1456–1461.
          pmc: PMC3165212pubmed: 21752949
        16. Malik P, Khurana SK, Dwivedi SK. Re-emergence of glanders in India: report of Maharashtra state.. Indian J. Microbiol. 2010 50:345–348.
          pmc: PMC3450061pubmed: 23100851
        17. Mukhopadhyay S. High-redundancy draft sequencing of 15 clinical and environmental Burkholderia strains.. J. Bacteriol. 2010 192:6313–6314.
          pmc: PMC2981208pubmed: 20870763
        18. Naureen A. Antimicrobial susceptibility of 41 Burkholderia mallei isolates from spontaneous outbreaks of equine glanders in Punjab, Pakistan.. J. Equine Vet. Sci. 2010 30:134–141.
        19. Naureen A, Saqib M, Muhammad G, Hussain MH, Asi MN. Comparative evaluation of Rose Bengal plate agglutination test, mallein test and some conventional serological tests for diagnosis of equine glanders.. J. Vet. Diagn. Invest. 2007 19:362–367.
          pubmed: 17609344
        20. Neubauer H, Meyer H, Finke EJ. Human glanders.. Rev. Int. Serv. Sante Forces Armees 1997 70:258–265.
        21. Neubauer H. Serodiagnosis of Burkholderia mallei infections in horses: state-of-the-art and perspectives.. J. Vet. Med. B 2005 52:201–205.
          pubmed: 16115091
        22. OIE. Glanders, chapter 2.5.11.. Manual of diagnostic tests and vaccines for terrestrial animals 2010 OIE, Paris, France.
        23. Parthasarathy N, DeShazer D, England M, Waag DM. Polysaccharide microarray technology for the detection of Burkholderia pseudomallei and Burkholderia mallei antibodies.. Diagn. Microbiol. Infect. Dis. 2006 56:329–332.
          pmc: PMC7127370pubmed: 16765554
        24. Pearson T. Phylogeographic reconstruction of a bacterial species with high levels of lateral gene transfer.. BMC Biol. 2009 7:78.
          doi: 10.1186/1741-7007-7-78pmc: PMC2784454pubmed: 19922616google scholar: lookup
        25. Schlater LK. Glanders.. Current therapy in equine medicine 1992 W. B. Saunders, St. Louis, MO.
        26. Sen GP, Singh G, Joshi TP. Comparative efficacy of serological tests in the diagnosis of glanders.. Indian Vet. J. 1968 45:286.
          pubmed: 4970533
        27. Sprague LD, Neubauer H. Melioidosis in animals: a review on epizootiology, diagnosis and clinical presentation.. J. Vet. Med. B 2004 51:305–320.
          pubmed: 15525357
        28. Sprague LD. A possible pitfall in the identification of Burkholderia mallei using molecular identification systems based on the sequence of the flagellin fliC gene.. FEMS Immunol. Med. Microbiol. 2002 34:231–234.
          pubmed: 12423776
        29. Sprague LD. Prevalence-dependent use of serological tests for diagnosing glanders in horses.. BMC Vet. Res. 2009 5:32.
          pmc: PMC2745380pubmed: 19723336
        30. Srinivasan A. Glanders in a military research microbiologist.. N. Engl. J. Med. 2001 345:256–258.
          pubmed: 11474663
        31. Steele JH. Bacterial, rickettsial and mycotic diseases, p 339–351. CRC handbook series in zoonoses, section A, vol I 1980 CRC Press LLC, Boca Raton, FL.
        32. Towbin H, Staehelin T, Gordin J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.. Proc. Natl. Acad. Sci. U. S. A. 1979 76:4350–4354.
          pmc: PMC411572pubmed: 388439
        33. Verma RD, Sharma JK, Venkateswaran KS, Batra HV. Development of an avidin-biotin dot enzyme-linked immunosorbent assay and its comparison with other serological tests for diagnosis of glanders in equines.. Vet. Microbiol. 1990 25:77–85.
          pubmed: 2247938

        Citations

        This article has been cited 9 times.
        1. Ichikawa Y, Iinuma Y, Okagawa T, Shimbo R, Enkhtuul B, Khurtsbaatar O, Kinoshita Y, Niwa H, Aoshima K, Kobayashi A, Batbaatar V, Ohashi K, Kimura T. Comparison of immunogenicity of 17 Burkholderia mallei antigens and whole cell lysate using indirect ELISA. J Vet Med Sci 2025 Apr 1;87(4):394-401.
          doi: 10.1292/jvms.25-0039pubmed: 40044168google scholar: lookup
        2. Abnaroodheleh F, Mosavari N, Pourbakhsh SA, Tadayon K, Jamshidian M. Identification of Burkholderia mallei Isolates with Polymerase Chain Reaction-Restriction Fragment Length Polymorphism. Arch Razi Inst 2023 Aug;78(4):1305-1312.
          doi: 10.32592/ARI.2023.78.4.1305pubmed: 38226390google scholar: lookup
        3. Charron P, Gao R, Chmara J, Hoover E, Nadin-Davis S, Chauvin D, Hazelwood J, Makondo K, Duceppe MO, Kang M. Influence of genomic variations on glanders serodiagnostic antigens using integrative genomic and transcriptomic approaches. Front Vet Sci 2023;10:1217135.
          doi: 10.3389/fvets.2023.1217135pubmed: 38125681google scholar: lookup
        4. Knox A, Zerna G, Beddoe T. Current and Future Advances in the Detection and Surveillance of Biosecurity-Relevant Equine Bacterial Diseases Using Loop-Mediated Isothermal Amplification (LAMP). Animals (Basel) 2023 Aug 18;13(16).
          doi: 10.3390/ani13162663pubmed: 37627456google scholar: lookup
        5. Elschner MC, Melzer F, Singha H, Muhammad S, Gardner I, Neubauer H. Validation of a Commercial Glanders ELISA as an Alternative to the CFT in International Trade of Equidae. Front Vet Sci 2021;8:628389.
          doi: 10.3389/fvets.2021.628389pubmed: 33665218google scholar: lookup
        6. Erdemsurakh O, Ochirbat K, Gombosuren U, Tserendorj B, Purevdorj B, Vanaabaatar B, Aoshima K, Kobayashi A, Kimura T. Seroprevalence of equine glanders in horses in the central and eastern parts of Mongolia. J Vet Med Sci 2020 Sep 24;82(9):1247-1252.
          doi: 10.1292/jvms.20-0219pubmed: 32641602google scholar: lookup
        7. Yazdansetad S, Mosavari N, Tadayon K, Mehregan I. Development of an immunoblotting assay for serodiagnosis of Burkholderia mallei infection: the whole-cell proteome-based paradigm. Iran J Microbiol 2019 Jun;11(3):232-238.
          pubmed: 31523407
        8. Lowe CW, Satterfield BA, Nelson DB, Thiriot JD, Heder MJ, March JK, Drake DS, Lew CS, Bunnell AJ, Moore ES, O'Neill KL, Robison RA. A Quadruplex Real-Time PCR Assay for the Rapid Detection and Differentiation of the Most Relevant Members of the B. pseudomallei Complex: B. mallei, B. pseudomallei, and B. thailandensis. PLoS One 2016;11(10):e0164006.
          doi: 10.1371/journal.pone.0164006pubmed: 27736903google scholar: lookup
        9. Singha H, Malik P, Goyal SK, Khurana SK, Mukhopadhyay C, Eshwara VK, Singh RK. Optimization and validation of indirect ELISA using truncated TssB protein for the serodiagnosis of glanders amongst equines. ScientificWorldJournal 2014;2014:469407.
          doi: 10.1155/2014/469407pubmed: 24672321google scholar: lookup
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