Detection of CWD prions in urine and saliva of deer by transgenic mouse bioassay.
Detection of CWD prions in urine and saliva of deer by transgenic mouse bioassay.
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DOI：
10.1371/journal.pone.0004848
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发表时间：
2009
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：
Haley NJ;Seelig DM;Zabel MD;Telling GC;Hoover EA
慢性消耗性疾病(CWD)是一种感染圈养和自由放养的鹿类(如鹿、麋鹿和驼鹿)的普恩病毒病。尽管体液被认为起着重要作用，但CWD的传播机制尚不清楚。在这里，我们报告了患有慢性消耗性疾病(CWD)的鹿的尿液和唾液中存在传染性普恩病毒。在表达Cervid PrP基因的转基因小鼠(TG[CerPrP]小鼠)中，通过浓缩、透析后的尿液和唾液的生物测定来检测Prion的感染性。用蛋白质错折叠循环扩增法(PMCA)检测混合尿和浓缩尿中PrPCWD。体液中异常蛋白含量很低，表现为：传统方法(免疫印迹、酶联免疫吸附试验)检测不到PrPCWD，接种的TG(CerPrP)小鼠潜伏期延长，TSE不完全性发作(9只尿液接种小鼠中2只373±3天，9只唾液接种小鼠8只342±109天)。这些发现有助于扩大我们对CWD Prion脱落和传播的理解，并预示着在其他Prion感染的体液中检测到感染性Prion。
Chronic wasting disease (CWD) is a prion disease affecting captive and free-ranging cervids (e.g. deer, elk, and moose). The mechanisms of CWD transmission are poorly understood, though bodily fluids are thought to play an important role. Here we report the presence of infectious prions in the urine and saliva of deer with chronic wasting disease (CWD). Prion infectivity was detected by bioassay of concentrated, dialyzed urine and saliva in transgenic mice expressing the cervid PrP gene (Tg[CerPrP] mice). In addition, PrPCWD was detected in pooled and concentrated urine by protein misfolding cyclic amplification (PMCA). The concentration of abnormal prion protein in bodily fluids was very low, as indicated by: undetectable PrPCWD levels by traditional assays (western blot, ELISA) and prolonged incubation periods and incomplete TSE attack rates in inoculated Tg(CerPrP) mice (373±3days in 2 of 9 urine-inoculated mice and 342±109 days in 8 of 9 saliva-inoculated mice). These findings help extend our understanding of CWD prion shedding and transmission and portend the detection of infectious prions in body fluids in other prion infections.
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