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首页> 外文期刊>BMC Biotechnology >Development of a diagnostic method for neosporosis in cattle using recombinant Neospora caninum proteins
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Development of a diagnostic method for neosporosis in cattle using recombinant Neospora caninum proteins

机译：利用重组犬新孢子虫蛋白开发牛新孢子虫病诊断方法

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Background Neosporosis is an infectious disease primarily of cattle and dogs, caused by intracellular parasite, Neospora caninum . Neosporosis appears to be a major cause of abortion in dairy cattle worldwide and causes to huge economic loss to dairy industry. Results Recombinant surface associated antigen 1 (NcSAG1), NcSAG1 related sequence 2 (NcSRS2) and the dense granule antigen 2 (NcGRA2) of N. caninum were expressed either in silkworm or in Escherichia coli and purified. The purified recombinant proteins bound to the N. caninum -specific antibodies in serum samples from infected cattle as revealed by an enzyme-linked immunosorbent assay ( ELISA ). By co-immobilizing these recombinant proteins, a novel indirect ELISA was developed for detection of neosporosis. With the use of 32 serum samples, comprising 12 positive serum samples and 20 negative serum samples, the sensitivity and specificity of the assay were found to be 91.7 and 100%, respectively. Seventy-two serum samples from dairy farms were also tested and one was diagnosed with neosporasis with both this method and a commercial assay. Conclusions A diagnostic method employing recombinant proteins of N. caninum was developed. The method showed high sensitivity and specificity. Diagnostic test with field serum samples suggested its applicability to the practical diagnosis of neosporosis.
机译：背景新孢子虫病是一种主要由牛和狗引起的传染病，由细胞内寄生虫犬新孢子虫引起。新孢子虫病似乎是全世界奶牛流产的主要原因，并给奶业造成巨大的经济损失。结果犬新孢子虫的重组表面相关抗原1（NcSAG1），NcSAG1相关序列2（NcSRS2）和致密颗粒抗原2（NcGRA2）在家蚕或大肠杆菌中表达并纯化。通过酶联免疫吸附测定（ELISA）揭示，纯化的重组蛋白与感染牛血清样品中的犬新孢子虫特异性抗体结合。通过共同固定这些重组蛋白，开发了一种新的间接ELISA用于检测新孢子虫病。通过使用32个血清样品（包括12个阳性血清样品和20个阴性血清样品），测定的灵敏度和特异性分别为91.7和100％。还对来自奶牛场的72个血清样品进行了测试，并通过该方法和商业化验方法将其中一个诊断为新孢子虫病。结论建立了犬新孢子虫重组蛋白诊断方法。该方法显示出高灵敏度和特异性。野外血清样品的诊断测试表明其可用于新孢子虫病的实际诊断。

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