Retrotransposon assay for tick host bloodmeal identification

用于蜱宿主血粉鉴定的逆转录转座子测定

• 批准号: 10211119
• 负责人: Sam R Telford
• 金额: $ 40.78万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-06 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10211119
• 关键词: American; Animals; Archives; Babesia microti; Babesiosis; Biological Assay; Birds; Black-legged Tick; Blood; Blood specimen; Borrelia burgdorferi; Borrelia miyamotoi; Canis familiaris; Collection; Complement; Costs and Benefits; DNA; Deer Tick; Development; Disease; Ecology; Encephalitis; Entomology; Event; Forensic Sciences; Genetic; Genome; Genomics; Goals; Gossypium; Hamsters; Human; Infection; Intervention; Literature; Lyme Disease; Mammals; Methods; Modification; Mus; New England; Nymph; Oral; Permethrin; Peromyscus; Public Health; Publishing; Recombinant DNA; Recording of previous events; Reproducibility; Retrotransposon; Risk; Sampling; Seasons; Site; Solid; Source; Testing; Tick-Borne Infections; Ticks; Time; Tube; United States; Vaccination; Vaccines; Virus; Virus Integration; White-Footed Mouse; Zoonoses; acaricide; expectation; field study; genome analysis; indexing; interest; microbial; spatiotemporal; tick feeding; tick transmission; tick-borne; tool; transmission process; vector; vertebrate genome

Project Summary/Abstract. The public health burden due to Lyme disease continues to expand in the eastern United States. New interventions to reduce risk are needed to complement well-known methods such as acaricidal applications or personal protection. Reservoir targeted approaches such as oral vaccination, feed through acaricides or genetic modifications are currently under development or are being field tested. These newer strategies focus on the white footed mouse, Peromyscus leucopus, which is widely regarded as the main host for subadult deer ticks (Ixodes dammini or I. scapularis), the vector for Lyme disease and diverse other zoonotic infections in the northeastern U.S. Indeed, this mouse has been incriminated as the main northeastern U.S. reservoir host for the agent of Lyme disease (Borrelia burgdorferi) as well as for that of human babesiosis (Babesia microti) and deer tick virus encephalitis. However, the evidence that P. leucopus is the main host for subadult deer ticks is largely descriptive using indirect extrapolations and comprise limited observations from a small number of sites. The ideal means of incriminating a reservoir host is to determine the source of the bloodmeal that served to infect the vector but assays that have been described to date for ticks have been poorly reproducible or insensitive. Our overarching goal for this proposed project is to provide a means to better understand natural variability of the relative contribution of other host species on the transmission of the agent of Lyme disease. This is required to evaluate the costs and benefits of mouse targeted interventions. We have determined that host retrotransposons may be targeted by PCR assays and provide specific signatures of the bloodmeal taken by a tick in its previous stage, and now seek to refine and validate this mode of analysis. Accordingly, we seek to (1) develop and validate a sensitive, specific, simple and reproducible retrotransposon assay for identifying the remnants of host bloodmeals in host-seeking nymphal deer ticks; (2) use this assay to determine whether the Peromyscus blood index (PBI) may vary across sites and within sites over time; and (3) experimentally confirm the PBI by undertaking interventions targeted to mice and assaying host seeking nymphal deer ticks thereafter to determine whether there has been a shift in hosts. Although our immediate goal is to provide a solid rationale for mouse targeted interventions against Lyme disease, the assay itself could be adapted to better understand the mode of perpetuation of diverse other tick-borne zoonoses.

项目摘要/摘要。 在美国东部，由于莱姆病造成的公共卫生负担继续扩大。 需要新的干预措施来降低风险，以补充诸如知名方法，例如 杀菌应用或个人保护。储层有针对性的方法，例如口服 目前正在开发疫苗接种，通过急经剂或遗传修饰的进食 或正在对现场测试。这些较新的策略专注于白色脚的鼠标Peromyscus 白菜，被广泛认为是亚鹿tick虫的主要主机（ixodes dammini或 I. scapularis），莱姆病的载体和其他人畜共患感染。 确实，美国东北部，这只老鼠被罪名为美国东北部的主要 莱姆病代理（Borrelia burgdorferi）以及人类的水库主机 巴贝斯病（Babesia microti）和鹿tick病毒脑炎。但是，证明P。 白牙是亚成年鹿tick的主要主机，在很大程度上是使用间接的 外推和包括少数站点的有限观察结果。理想 犯罪储存宿主的手段是确定所服务的血液的来源 感染矢量，但迄今为止描述的tick的测定很差 可再现或不敏感。我们对这个拟议项目的总体目标是提供 更好地了解其他宿主物种相对贡献的自然变异性 关于莱姆病药物的传播。这是评估成本和 鼠标靶向干预措施的好处。我们已经确定主机逆转座子可能 由PCR分析对准，并提供特定的签名 上一个阶段，现在试图完善和验证这种分析方式。因此，我们寻求 （1）开发和验证一种敏感，特异性，简单且可重复的逆转录座子测定 用于识别在寻求宿主的若虫鹿中的宿主血液的残留物； （2）使用此 测定确定peromyscus血液指数（PBI）是否可能在各个地点和内部有所不同 随着时间的流逝； （3）通过实现针对的干预措施，实验确认PBI 此后，向小鼠和分析宿主寻求若虫鹿的tick虫来确定是否有 是主机的转变。尽管我们的近期目标是为鼠标提供坚实的基本原理 针对莱姆病的有针对性干预措施，该测定本身可以改善 了解其他不同tick传播的人畜共患病的延续方式。