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

BMC Veterinary Research

Jenner Karlisson Pimenta dos Reis¹  Rômulo Cerqueira Leite¹  Paulo Rodrigues Lopes Filho³  Evandro dos Reis Machado³  Marcilia Maria Alves de Souza²  Vania Lucia de Assis Santana²  Mariana Lázaro Sales⁴  Lívia de Lima Orzil⁴  Antônio Augusto Fonseca⁴  Rodrigo Mauro Ramos⁴  Maurício Baltazar de Carvalho Filho⁴ 

[1] Departamento de Medicina Veterinária Preventiva, Escola de Veterinária - Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, Pampulha, Belo Horizonte, 31270-901, MG, Brazil;Laboratório Nacional Agropecuário em Pernambuco, Recife, Brazil;Instituto Mineiro de Agropecuária, Belo Horizonte, Brazil;Laboratório Nacional Agropecuário em Minas Gerais, Pedro Leopoldo, Brazil

关键词: Purification;    Diagnosis;    Antigen;    Mallein;    Glanders;    Equine;   

Others  :  1119733
DOI  :  10.1186/1746-6148-8-154

 received in 2012-03-22, accepted in 2012-08-28,  发布年份 2012

【 摘 要 】

Background

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.

Conclusion

Some of the TFF-purified malleins showed considerable potential to be used as an auxiliary test in the diagnosis of glanders.

【 授权许可】

   
2012 Filho et al.; licensee BioMed Central Ltd.

【 图 表 】

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