Infection and prevalence of a new segmented flavivirus in the United States

总结/摘要 在解旋酶中与原型黄病毒具有系统发生相关性的节段RNA病毒 (NS3)和聚合酶（NS5）蛋白最近从蜱，蚊子和脊椎动物，包括 患有发热性疾病的人第一批黄病毒样节段病毒（FLSV）是从蜱虫中分离出来的， 荆门蜱病毒是中国荆门地区的一种蜱类病毒，因此被命名为荆门蜱病毒。随后，高度多样化的 在非洲、欧洲和南美洲发现了FLSV变异体。然而，没有FLSV感染的证据， 在包括美国在内的北美也有报道。此外，最近的几项研究， 基于宏基因组学的蜱中病毒组的鉴定未能鉴定FLSV感染。我们在描述 北美最常见的野生鼠种- 美国，当我们在宾夕法尼亚州林地的几只鹿鼠中鉴定出高度多样的FLSV时（FLSV- PN）。我们获得了FLSV-PN基因组所有五个片段的部分序列，并开发了血清学检测试剂盒。 使用荧光素酶免疫沉淀系统的测定。我们的血清学数据表明，FLSV-PN或其相关 变异体具有广泛的宿主范围并感染几种经济上重要的农场和家畜。最 重要的是，我们在有蜱叮咬史和莱姆病史的人中发现了FLSV-PN感染的有力证据 疾病我们的探索性/开发性提案的长期目标是确定发病机制和健康状况 FLSV-PN感染的相关性。开发这种新病毒的分子试剂将使我们能够测试我们的 假设几种遗传多样的FLSV变体在美国流行，这些病毒 主要通过蜱传播给人类和动物。具体目标-1是获得完整的 几种不同的FLSV变体的基因组。我们的初步数据表明，FLSV-PN基因组由 5个RNA片段在3 '端被多聚腺苷酸化。FLSV-PN全基因组测序 开发敏感和特异性诊断测定以研究感染流行率所必需的， 疾病协会。具体目标-2是确定FLSV在人类中的感染率， 动物和蜱虫这些研究将得到来自以下临床研究的样本和数据的帮助： 莱姆病在NIAID，NIH（支持信附后）。拟议的目标代表着朝着以下目标迈出的关键步骤： FLSV-PN的详细分子和生物学特征，包括反向遗传学的开发 系统、复制子系统、细胞培养和动物模型。虽然FLSV感染的健康相关性是 目前尚不清楚，拟议的研究将告知这一知识，以及开发试剂， 如有必要，防止这种新病毒和相关疾病的传播。

Summary/abstract Segmented RNA viruses that share phylogenetic relatedness with the prototypic Flaviviruses in their helicase (NS3) and polymerase (NS5) proteins were recently identified from ticks, mosquitoes, and vertebrates, including humans suffering with febrile illness. The first Flavi-like segmented viruses (FLSV) were isolated from ticks and animals in the Jingmen province of China and were thus named Jingmen Tick Virus. Subsequently, highly diverse FLSV variants were found in Africa, Europe and South America. However, no evidence of FLSV infection has been reported in North America, including the United States. Moreover, several recent studies that used metagenomics-based identification of the virome in ticks failed to identify FLSV infection. We were characterizing the virome of free-ranging mice of the genus Peromyscus, the most common wild mouse species of the North America, when we identified a highly diverse FLSV in several deer mice from woodlands in Pennsylvania (FLSV- PN). We acquired partial sequences of all five segments of the FLSV-PN genome and developed serological assays using Luciferase immunoprecipitation systems. Our serological data suggest that FLSV-PN or its related variants have a wide host range and infect several economically important farm and domestic animals. Most importantly, we found strong evidence of FLSV-PN infection in humans with a history of tick-bites and Lyme disease. The long-term goal of our exploratory/developmental proposal is to define the pathogenesis and health relevance of FLSV-PN infection. Development of molecular reagents for this new virus will allow us to test our hypotheses that several genetically diverse FLSV variants are circulating in the United States, and these viruses are predominantly transmitted by ticks to humans and animals. Specific aim-1 is to acquire the complete genomes of several diverse FLSV variants. Our preliminary data suggest that FLSV-PN genome is comprised of five RNA segments that are polyadenylated at the 3’ end. Complete genome sequencing of FLSV-PN is necessary for the development of sensitive and specific diagnostic assays to study infection prevalence and disease associations. Specific aim-2 is to determine the infection prevalence of FLSV in humans, domesticated animals and ticks. These studies will be aided by samples and data available from clinical research studies of Lyme disease at NIAID, NIH (letter of support attached). The proposed aims represent crucial steps toward detailed molecular and biological characterization of FLSV-PN, including the development of a reverse genetics system, replicon systems, cell culture and animal models. Although the health relevance of FLSV infections is currently not known, the proposed studies will inform this knowledge as well as develop the reagents for preventing the spread of this new virus and associated diseases, if necessary.