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BMC veterinary research2012; 8; 154; doi: 10.1186/1746-6148-8-154

Development and validation of a method for purification of mallein for the diagnosis of glanders in equines.

Abstract: The allergic test of mallein is one of the most frequently used tests, together with the Complement Fixation Test (CFT), for the diagnosis of glanders in endemic areas. Mallein, a purified protein derivative (PPD), is produced similarly to PPD tuberculin and the end product is a primarily proteic antigen, which is only poorly purified. The immuno-allergic activity of mallein is believed to be due to a high molecular weight group of proteins present in the antigen. To improve the quality of the antigen, in terms of sensitivity and specificity, a new method of mallein production was developed, in which purification was accomplished by ultrafiltration in a Tangential Flow Filtration system (TFF). Results: The TFF methodology efficiently separated the high and low molecular weight protein groups of mallein. The five TFF-purified malleins, produced from Burkholderia mallei strains isolated from clinical cases of glanders in Brazil, proved to be more potent than standard mallein in the induction of an allergic reaction in sensitized animals. Regarding specificity, two of the purified malleins were equivalent to the standard and three were less specific. Conclusions: Some of the TFF-purified malleins showed considerable potential to be used as an auxiliary test in the diagnosis of glanders.
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Publication Date: 2012-09-02 PubMed ID: 22937975PubMed Central: PMC3514244DOI: 10.1186/1746-6148-8-154Google Scholar: Lookup
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Summary

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This research has developed and tested a new method for producing mallein, a protein used in diagnosing a bacterial infection in horses known as glanders. The new production method successfully purified the proteins in mallein and showed potential for being a more effective testing agent, although further refinement is needed to improve its specificity.

Understanding Glanders and Mallein

  • The focus of this study is glanders, a bacterial infection primarily affecting horses but also occasionally humans and other mammals.
  • Glanders is typically diagnosed using an allergic test that involves mallein, a purified protein derivative (PPD).
  • The quality and purification of mallein protein can greatly affect the accuracy of glanders diagnosis.
  • The operation of mallein in diagnosis is thought to be tied to a high molecular weight group of proteins present in the antigen.
  • However, the standard purification process for mallein only mildly purifies the proteic antigen, resulting in a need for improved purification methods.

Purification Process Development

  • To improve the quality of the mallein antigen, a new method of mallein production was developed.
  • In this new method, purification was accomplished using ultrafiltration in a Tangential Flow Filtration (TFF) system.
  • The TFF method effectively separated the high and low molecular weight protein groups of mallein.

Testing the Purified Malleins

  • The researchers then tested five TFF-purified mallein samples, all produced from Burkholderia mallei strains isolated from real-life cases of glanders in Brazil.
  • The results showed that the purified malleins were more potent in inducing an allergic reaction in sensitized animals, suggesting improved sensitivity compared to the standard mallein.
  • However, in terms of specificity, only two of the purified malleins matched the standard, while the other three were less specific.
  • This indicates that while some of the TFF-purified malleins show promise as a potentially more effective diagnostic test for glanders, further refinement may be necessary to address issues of specificity.

Cite This Article

APA
de Carvalho Filho MB, Ramos RM, Fonseca AA, de Lima Orzil L, Sales ML, de Assis Santana VL, de Souza MM, Dos Reis Machado E, Filho PR, Leite RC, Dos Reis JK. (2012). Development and validation of a method for purification of mallein for the diagnosis of glanders in equines. BMC Vet Res, 8, 154. https://doi.org/10.1186/1746-6148-8-154

Publication

BMC veterinary research
ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 8
Pages: 154

Researcher Affiliations

de Carvalho Filho, Maurício Baltazar
  • Departamento de Medicina Veterinária Preventiva, Escola de Veterinária - Universidade Federal de Minas Gerais, Av, Antonio Carlos 6627, Pampulha, Belo Horizonte, 31270-901, MG, Brazil.
Ramos, Rodrigo Mauro
    Fonseca, Antônio Augusto
      de Lima Orzil, Lívia
        Sales, Mariana Lázaro
          de Assis Santana, Vania Lucia
            de Souza, Marcilia Maria Alves
              Dos Reis Machado, Evandro
                Filho, Paulo Rodrigues Lopes
                  Leite, Rômulo Cerqueira
                    Dos Reis, Jenner Karlisson Pimenta

                      MeSH Terms

                      • Animals
                      • Antibodies, Bacterial / blood
                      • Antigens, Bacterial / immunology
                      • Burkholderia mallei / classification
                      • Burkholderia mallei / metabolism
                      • Complement Fixation Tests / veterinary
                      • Glanders / diagnosis
                      • Guinea Pigs
                      • Horses
                      • Immunologic Tests / veterinary
                      • Reproducibility of Results
                      • Sensitivity and Specificity

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                      Citations

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