Rapid antemortem detection of CWD prions in deer saliva.
Rapid antemortem detection of CWD prions in deer saliva.
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DOI：
10.1371/journal.pone.0074377
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Hoover EA
中科院分区：
文献类型：
--
作者：
Henderson DM;Manca M;Haley NJ;Denkers ND;Nalls AV;Mathiason CK;Caughey B;Hoover EA
慢性消耗性疾病（CWD）是一种通过朊病毒有效传播的鹿病，目前已在美国22个州、加拿大2个省和韩国发现。慢性消耗病的一个标志是唾液中传染性朊病毒的脱落，如鹿的生物测定所示。同样清楚的是，朊病毒在唾液、血液、尿液和粪便中的浓度远低于神经系统或淋巴组织中的浓度。体液和排泄物中CWD（和其他）朊病毒的快速体外检测一直存在问题，这是由于直接测定（蛋白质印迹，ELISA）的灵敏度限制以及这些复杂生物材料中存在阻碍检测的抑制剂。在这里，我们使用实时震动诱导转换（RT-QuIC），以证明慢性消耗病朊病毒在稀释和朊病毒富集唾液样本从无症状和有症状的白尾鹿。在24份暴露于CWD的白尾鹿稀释唾液样本中，有14份（58.3%）检测到CWD朊病毒，其中包括14只无症状动物中的9只（64.2%）。此外，磷钨酸富集增强了RT-QuIC测定的灵敏度，使得能够在上述唾液样品中的24个中的19个（79.1%）中进行检测。在Tg[CerPrP]小鼠中的生物测定证实了在如此检查的2个RT-QuIC阳性唾液样品中的2个中存在感染性朊病毒。所描述的改良RT-QuIC分析代表了复杂生物液体、排泄物或环境样品中朊病毒的非侵入性、快速死前检测，以及探索朊病毒贩运、外周化和传播的工具。
Chronic wasting disease (CWD) is an efficiently transmitted prion disease of cervids, now identified in 22 United States, 2 Canadian provinces and Korea. One hallmark of CWD is the shedding of infectious prions in saliva, as demonstrated by bioassay in deer. It is also clear that the concentration of prions in saliva, blood, urine and feces is much lower than in the nervous system or lymphoid tissues. Rapid in vitro detection of CWD (and other) prions in body fluids and excreta has been problematic due to the sensitivity limits of direct assays (western blotting, ELISA) and the presence of inhibitors in these complex biological materials that hamper detection. Here we use real-time quaking induced conversion (RT-QuIC) to demonstrate CWD prions in both diluted and prion-enriched saliva samples from asymptomatic and symptomatic white-tailed deer. CWD prions were detected in 14 of 24 (58.3%) diluted saliva samples from CWD-exposed white-tailed deer, including 9 of 14 asymptomatic animals (64.2%). In addition, a phosphotungstic acid enrichment enhanced the RT-QuIC assay sensitivity, enabling detection in 19 of 24 (79.1%) of the above saliva samples. Bioassay in Tg[CerPrP] mice confirmed the presence of infectious prions in 2 of 2 RT-QuIC-positive saliva samples so examined. The modified RT-QuIC analysis described represents a non-invasive, rapid ante-mortem detection of prions in complex biologic fluids, excreta, or environmental samples as well as a tool for exploring prion trafficking, peripheralization, and dissemination.
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影响因子：
3.7
作者：
Haley NJ;Seelig DM;Zabel MD;Telling GC;Hoover EA
通讯作者：
Hoover EA
影响因子：
3.5
作者：
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通讯作者：
Soto, Claudio
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6.7
作者：
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通讯作者：
Prusiner SB
影响因子：
5.4
作者：
Heisey, Dennis M.;Mickelsen, Natalie A.;Barr, Daniel J.
通讯作者：
Barr, Daniel J.
影响因子：
9.9
作者：
BENDHEIM, PE;BROWN, HR;BOLTON, DC
通讯作者：
BOLTON, DC