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Expression and Immunogenicity of Recombinant Immunoreactive Surface Protein 2 of Anaplasma phagocytophilum

机译:吞噬嗜浆细胞重组免疫反应表面蛋白2的表达及免疫原性

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摘要

Human granulocytic anaplasmosis (HGA), caused by Anaplasma phagocytophilum, is an emerging tick-borne zoonotic disease throughout the world. The first HGA cases in China were documented in 2008, and the greatest challenge posed by the disease is rapid and accurate diagnosis during the acute phage of illness. In this study, we successfully cloned and expressed an A. phagocytophilum immunoreactive surface protein (major surface protein 2 [MSP2]) and demonstrated that this recombinant protein has natural immunogenicity by Western blotting and enzyme-linked immunosorbent assay (ELISA) using human HGA-positive sera and reference rabbit HGA-positive sera. The rabbit antisera against the recombinant protein also reacted actively with the natural antigen of A. phagocytophilum by immunofluorescence assay (IFA). No cross-reaction was observed between the recombinant protein and rabbit antisera against 10 common members of the order Rickettsiales by ELISA when the sera were diluted more than 1:100. We concluded that the recombinant MSP2 protein exhibited excellent antigenicity and specificity, results that should lay the foundation for the development of a simple and rapid diagnostic reagent and a vaccination for anaplasmosis.
机译:由吞噬性无浆膜炎引起的人类粒细胞无性病(HGA)是全世界新兴的由tick传播的人畜共患病。中国首例HGA病例记录于2008年,该疾病带来的最大挑战是在急性噬菌体疾病期间如何快速准确地进行诊断。在这项研究中,我们成功克隆并表达了嗜A.phagocytophilum免疫反应性表面蛋白(主要表面蛋白2 [MSP2]),并通过Western印迹和酶联免疫吸附测定(ELISA)使用人类HGA-阳性血清和参考兔HGA阳性血清。通过免疫荧光测定法(IFA),针对重组蛋白的兔抗血清也与吞噬曲霉的天然抗原活跃反应。当血清以超过1:100的比例稀释时,重组蛋白与兔抗血清之间没有针对立克次体的10个常见成员的交叉反应。我们得出的结论是,重组MSP2蛋白表现出优异的抗原性和特异性,这一结果应为开发一种简单,快速的诊断试剂和进行无病菌接种奠定基础。

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