PMCA Detection of CWD Infection in Cervid and Non-Cervid Species

PMCA 检测鹿科动物和非鹿科动物的 CWD 感染

• 批准号: 7779776
• 负责人: EDWARD Arthur HOOVER
• 金额: $ 31.02万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7779776
• 关键词: Animals; Asians; Biological; Biological Assay; Blood; Body Fluids; Bovine Spongiform Encephalopathy; Brain; Carrier State; Chronic Wasting Disease; Colorado; Country; Deer; Detection; Diagnosis; Enzyme-Linked Immunosorbent Assay; Epidemiologic Studies; Excretory function; Exposure to; Feces; Health; Human; Immunohistochemistry; In Vitro; Individual; Infection; Korea; Laboratories; Lateral; Life; Liquid substance; Lymphoid Tissue; Methodology; Mus; Muscle; Nature; Peripheral; PrP; PrPCWD; Predisposition; Prion Diseases; Prions; Protein Isoforms; Province; Relative (related person); Risk; Saliva; Sampling; Source; Technology; Testing; Tissues; Transgenic Mice; Urine; Western Blotting; Wyoming; base; cervid; disease transmission; improved; in vitro Assay; in vivo; insight; protein misfolding cyclic amplification; public health relevance; transmission process

DESCRIPTION (provided by applicant): Chronic wasting disease (CWD), an emergent, efficiently transmitted, fatal prion disease of cervids (deer, elk, moose), is now identified in 15 states, 2 Canadian provinces, and Korea. We have found that infectious prions capable of transmitting CWD are present in the saliva, blood, and urine of CWD-infected deer, potentially explaining the lateral transmission of the disease. The presence of CWD prions in saliva and blood confirms prion infectivity in biological fluids and provides realistic hope for antemortem detection via a more sensitive in vitro assay, which forms the basis for this application. Direct detection of the abnormal CWD prion protein (PrPRES/CWD) in body fluids or excreta of deer has not been possible using contemporary assay methodologies (i.e. immunohistochemistry, western blotting, ELISA). Here we propose to employ serial protein misfolding cyclic amplification (sPMCA) to amplify and detect PrPCWD in saliva, blood, urine, and feces of infected deer. Direct in vitro detection of PrPCWD would enable pre-symptomatic ante-mortem diagnosis, improve ecologic understanding of CWD, and potentially enable detection of prion carrier states and contamination in cervid products. Whether a non-cervid reservoir for CWD infection exists in nature remains unknown. Likewise, whether CWD represents a risk to human health is uncertain. To begin to assess the potential for trans-species transmission of CWD, we propose to use sPMCA as a surrogate to assess whether the normal prion protein from selected non-cervid species can be converted to the abnormal PrPCWD isoform after exposure to CWD prions. This information will be used to direct correlative in vivo inoculation and epidemiologic studies needed to assess the CWD species barrier, identify potential reservoir hosts in nature, and better estimate potential human or other species health risks posed by CWD. PUBLIC HEALTH RELEVANCE: Chronic wasting disease (CWD) is a rapidly spreading fatal prion disease of deer and elk, probably spread through saliva, and now present in 15 states. Whether CWD may represent a risk to human health analogous to mad cow disease is unknown. Proposed here is a new more powerful test to detect CWD prions in saliva, blood, and excretions of infected deer-something not possible with current technology yet vital to controlling the spread of CWD among deer and potentially other species.

描述（由申请人提供）：慢性消耗性疾病 (CWD) 是一种新出现的、有效传播的、致命的鹿科动物（鹿、麋鹿、驼鹿）朊病毒病，现已在 15 个州、加拿大 2 个省和韩国发现。我们发现，感染 CWD 的鹿的唾液、血液和尿液中存在能够传播 CWD 的传染性朊病毒，这可能解释了该疾病的横向传播。唾液和血液中 CWD 朊病毒的存在证实了生物体液中朊病毒的感染性，并为通过更灵敏的体外测定进行生前检测提供了现实的希望，这构成了该应用的基础。使用当代检测方法（即免疫组织化学、蛋白质印迹、ELISA）无法直接检测鹿体液或排泄物中的异常 CWD 朊病毒蛋白 (PrPRES/CWD)。在这里，我们建议采用系列蛋白错误折叠循环扩增 (sPMCA) 来扩增和检测受感染鹿的唾液、血液、尿液和粪便中的 PrPCWD。 PrPCWD 的直接体外检测将能够实现症状前的宰前诊断，提高对 CWD 的生态学理解，并有可能检测鹿产品中的朊病毒携带状态和污染。自然界中是否存在 CWD 感染的非鹿科动物宿主仍不清楚。同样，CWD 是否对人类健康构成风险也不确定。为了开始评估 CWD 跨物种传播的可能性，我们建议使用 sPMCA 作为替代物来评估来自选定非鹿科动物物种的正常朊病毒蛋白在暴露于 CWD 朊病毒后是否可以转化为异常的 PrPCWD 同工型。该信息将用于指导评估 CWD 物种屏障所需的相关体内接种和流行病学研究，识别自然界中潜在的储存宿主，并更好地估计 CWD 造成的潜在人类或其他物种健康风险。 公众健康相关性：慢性消耗性疾病 (CWD) 是一种在鹿和麋鹿中迅速传播的致命朊病毒疾病，可能通过唾液传播，目前存在于 15 个州。 CWD 是否对人类健康构成类似于疯牛病的风险尚不清楚。这里提出了一种新的更强大的测试，可以检测受感染鹿的唾液、血液和排泄物中的 CWD 朊病毒，这在现有技术下是不可能的，但对于控制 CWD 在鹿和其他潜在物种中的传播至关重要。