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

描述(申请人提供)：慢性消耗性疾病(CWD)是一种紧急的、有效传播的、致命的颈鹿(鹿、麋鹿、驼鹿)普里恩疾病，现已在15个州、加拿大2个省和韩国确诊。我们已经发现，感染CWD的鹿的唾液、血液和尿液中存在能够传播CWD的传染性普恩病毒，这可能解释了这种疾病的横向传播。唾液和血液中CWD病毒的存在证实了病毒在生物体液中的感染性，并为通过更灵敏的体外检测方法进行生前检测提供了现实的希望，这为这一应用奠定了基础。用现代检测方法(即免疫组织化学、免疫印迹、ELISA)无法直接检测到鹿体液或排泄物中异常的CWD病毒蛋白(PrPRES/CWD)。在这里，我们建议使用序列蛋白质错折叠循环扩增(SPMCA)来扩增和检测感染的鹿的唾液、血液、尿液和粪便中的PrPCWD。对PrPCWD的直接体外检测将能够进行症状前的死亡前诊断，提高对CWD的生态理解，并有可能检测PrPCWD的病毒携带者状态和Cervid产品中的污染。自然界中是否存在CWD感染的非宫颈宿主尚不清楚。同样，CWD是否对人类健康构成风险也是不确定的。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，评估来自选定的非鹿类物种的正常PrPCWD蛋白在暴露于CWD Prion后是否可以转化为异常的PrPCWD亚型。这些信息将被用于指导相关的体内接种和流行病学研究，以评估CWD的物种屏障，确定自然界中潜在的宿主，并更好地估计CWD对人类或其他物种构成的潜在健康风险。 与公共卫生相关：慢性消耗性疾病(CWD)是一种在鹿和麋鹿中迅速传播的致命的普恩病毒疾病，可能通过唾液传播，目前在15个州出现。CWD是否会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来检测CWD感染鹿的唾液、血液和排泄物中的Prion-这是目前的技术所不可能做到的，但对于控制CWD在鹿和潜在的其他物种中的传播至关重要。

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.