Phylogenetic analysis of Puumala virus subtype Bavaria, characterization and diagnostic use of its recombinant nucleocapsid protein
Phylogenetic analysis of Puumala virus subtype Bavaria, characterization and diagnostic use of its recombinant nucleocapsid protein
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DOI：
10.1007/s11262-011-0620-x
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发表时间：
2011-10-01
期刊：
影响因子：
1.6
通讯作者：
Essbauer, Sandra S.
中科院分区：
文献类型：
--
作者：
Mertens, Marc;Kindler, Eveline;Essbauer, Sandra S.
普马拉病毒（PUUV）是德国主要的汉坦病毒种类，引起大量轻至中度肾综合征出血热（HFRS）病例。在2004年德国东南部爆发的一次疫情中，一种名为巴伐利亚的新型puv亚型被确定为人类HFRS的病原体。本文通过研究2004年和2005年捕获的下巴伐利亚州田鼠（Myodes glareolus）的部分和几乎全部核衣壳(N)蛋白编码的小（S-）片段序列和部分中（M-）片段序列，对该PUUV菌株进行了分子表征。系统发育分析证实其分类为巴伐利亚亚型，该亚型进一步细分为四个地理集群。巴伐利亚菌株的整个N蛋白（含氨基末端六组氨酸标签）是在酵母酿酒酵母中产生的，与酵母表达的PUUV菌株Vranica/Hallnas的N蛋白相比，其对N特异性单克隆抗体的反应性略有不同。来自德国和芬兰不同地区的人血清终点滴定显示，不同重组蛋白的诊断价值只有非常微小的差异。基于新N抗原间接捕获和单克隆抗体捕获，建立了igg - elisa。通过使用德国和芬兰的血清面板，他们的验证显示出高灵敏度和特异性。总之，我们的研究表明，巴伐利亚PUUV菌株与其他PUUV菌株存在遗传差异，其N蛋白具有诊断应用的潜力。
Puumala virus (PUUV) is the predominant hantavirus species in Germany causing large numbers of mild to moderate cases of haemorrhagic fever with renal syndrome (HFRS). During an outbreak in South-East Germany in 2004 a novel PUUV subtype designated Bavaria was identified as the causative agent of HFRS in humans [1]. Here we present a molecular characterization of this PUUV strain by investigating novel partial and almost entire nucleocapsid (N) protein-encoding small (S-) segment sequences and partial medium (M-) segment sequences from bank voles (Myodes glareolus) trapped in Lower Bavaria during 2004 and 2005. Phylogenetic analyses confirmed their classification as subtype Bavaria, which is further subdivided into four geographical clusters. The entire N protein, harbouring an amino-terminal hexahistidine tag, of the Bavarian strain was produced in yeast Saccharomyces cerevisiae and showed a slightly different reactivity with N-specific monoclonal antibodies, compared to the yeast-expressed N protein of the PUUV strain Vranica/Hallnas. Endpoint titration of human sera from different parts of Germany and from Finland revealed only very slight differences in the diagnostic value of the different recombinant proteins. Based on the novel N antigen indirect and monoclonal antibody capture IgG-ELISAs were established. By using serum panels from Germany and Finland their validation demonstrated a high sensitivity and specificity. In summary, our investigations demonstrated the Bavarian PUUV strain to be genetically divergent from other PUUV strains and the potential of its N protein for diagnostic applications.