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

BMC Veterinary Research

Methodology Article

Rômulo Cerqueira Leite¹  Jenner Karlisson Pimenta dos Reis¹  Evandro dos Reis Machado²  Paulo Rodrigues Lopes Filho²  Mariana Lázaro Sales³  Maurício Baltazar de Carvalho Filho³  Rodrigo Mauro Ramos³  Lívia de Lima Orzil³  Antônio Augusto Fonseca³  Vania Lucia de Assis Santana⁴  Marcilia Maria Alves de Souza⁴ 

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

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

DOI  :  10.1186/1746-6148-8-154

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

来源: Springer

【 摘 要 】

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

【 授权许可】

Unknown   
© Filho et al.; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.