Development and validation of a method for purification of mallein for the diagnosis of glanders in equines
Development and validation of a method for purification of mallein for the diagnosis of glanders in equines
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DOI：
10.1186/1746-6148-8-154
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发表时间：
2012-09-02
影响因子：
2.6
通讯作者：
Pimenta dos Reis, Jenner Karlisson
中科院分区：
文献类型：
--
作者：
de Carvalho Filho, Mauricio Baltazar;Ramos, Rodrigo Mauro;Pimenta dos Reis, Jenner Karlisson
背景：马来蛋白过敏试验是与补体固定试验(CFT)一起用于诊断流行地区腺体的最常用的试验之一。马来蛋白(Mallein)是一种纯化蛋白衍生物(PPD)，其产生方式类似于PPD结核菌素，最终产物主要是一种蛋白质抗原，只是纯化程度较低。马来蛋白的免疫过敏活性被认为是由于抗原中存在的一组高分子量蛋白质所致。为了提高马来蛋白的质量，从灵敏度和特异度两方面考虑，建立了一种新的马来蛋白生产方法，即用切向流过滤系统(TFF)进行超滤纯化。从巴西临床病例中分离的马来伯克霍尔德氏菌产生的五种经TFF提纯的马来蛋白，在诱导致敏动物的过敏反应方面被证明比标准马来蛋白更有效。在特异性方面，2个纯化的锤蛋白与标准相当，3个低于标准。结论：部分纯化的锤蛋白有相当大的潜力作为腺体诊断的辅助试验。
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.