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Expression of recombinant Araraquara Hantavirus nucleoprotein in insect cells and its use as an antigen for immunodetection compared to the same antigen expressed in Escherichia coli

机译:与在大肠杆菌中表达的相同抗原相比,重组Araraquara汉坦病毒核蛋白在昆虫细胞中的表达及其作为免疫检测抗原的用途

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摘要

Abstract Background Antigens for Hantavirus serological tests have been produced using DNA recombinant technology for more than twenty years. Several different strategies have been used for that purpose. All of them avoid the risks and difficulties involved in multiplying Hantavirus in the laboratory. In Brazil, the Araraquara virus is one of the main causes of Hantavirus Cardio-Pulmonary Syndrome (HCPS). Methods In this investigation, we report the expression of the N protein of the Araraquara Hantavirus in a Baculovirus Expression System, the use of this protein in IgM and IgG ELISA and comparison with the same antigen generated in E. coli. Results The protein obtained, and purified in a nickel column, was effectively recognized by antibodies from confirmed HCPS patients. Comparison of the baculovirus generated antigen with the N protein produced in E. coli showed that both were equally effective in terms of sensitivity and specificity. Conclusions Our results therefore indicate that either of these proteins can be used in serological tests in Brazil.
机译:摘要背景二十多年来,使用DNA重组技术生产用于汉坦病毒血清学测试的抗原。为此目的使用了几种不同的策略。所有这些都避免了在实验室中繁殖汉坦病毒所带来的风险和困难。在巴西,Araraquara病毒是汉坦病毒心肺综合征(HCPS)的主要原因之一。方法在本研究中,我们报告了杆状病毒汉兰病毒N蛋白在杆状病毒表达系统中的表达,该蛋白在IgM和IgG ELISA中的使用以及与大肠杆菌中产生的相同抗原的比较。结果获得的蛋白质在镍柱中纯化,可被确诊的HCPS患者的抗体有效识别。杆状病毒产生的抗原与大肠杆菌中产生的N蛋白的比较表明,两者在敏感性和特异性方面都同样有效。结论因此,我们的结果表明,这些蛋白中的任何一种均可用于巴西的血清学检测。

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