Detection of sub-clinical CWD infection in conventional test-negative deer long after oral exposure to urine and feces from CWD+ deer.
Detection of sub-clinical CWD infection in conventional test-negative deer long after oral exposure to urine and feces from CWD+ deer.
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DOI：
10.1371/journal.pone.0007990
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发表时间：
2009-11-24
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;Mathiason CK;Zabel MD;Telling GC;Hoover EA
鹿科动物的慢性消耗性疾病（CWD）是一种以高水平传播性为特征的朊病毒疾病，其中体液和排泄物被认为发挥着重要作用。使用宫颈生物测定和已建立的 CWD 检测方法，我们之前已经在 CWD+ 供体的唾液和血液中鉴定出传染性朊病毒，但在尿液或粪便中未鉴定出传染性朊病毒。最近，我们通过 cervid PrP 转基因 (Tg[CerPrP]) 小鼠生物测定和系列蛋白错误折叠循环扩增 (sPMCA) 鉴定出鹿尿液中浓度非常低的 CWD 朊病毒。这一发现促使我们使用 sPMCA 进一步检查我们最初的鹿科动物生物测定实验。我们试图调查之前经口接触 CWD+ 来源的尿液和粪便的常规检测阴性鹿是否可能携带低水平 CWD 感染，而在 19 个月的观察期内并不明显。我们进一步尝试确定这些动物的外周 PrPCWD 分布。通过 sPMCA 和鹿转基因小鼠生物测定，与适当的组织匹配的阳性和阴性对照平行，重新分析来自常规测试阴性鹿的各种神经和淋巴组织中的 CWD 朊病毒。通过 sPMCA 和 Tg[CerPrP] 小鼠生物测定，在经口暴露的鹿的组织中检测到 PrPCWD；每次检测都显示 CWD 朊病毒的水平非常低，以前无法通过蛋白质印迹、ELISA 或 IHC 检测到。对个体组织进行的系列 PMCA 分析表明，在 5 只尿液/粪便暴露的鹿中，有 4 只仅 obex 呈阳性。 PrPCWD 从阳性对照鹿的淋巴组织和神经组织中扩增，但没有从阴性对照鹿的相同组织中扩增。在经口接触尿液和粪便的鹿中检测到亚临床感染 (1) 表明可能存在长期亚临床状态，需要两年以上的观察期才能检测 CWD 感染，(2) 说明 sPMCA 在诊断低水平朊病毒感染中的敏感性和特异性应用。根据这些结果，低剂量的朊病毒，例如在口服接触 CWD 感染的鹿的尿液和唾液后，可能会利用消化道和中枢神经系统之间的神经导管，绕过 LRS 的显着放大，正如在其他一些朊病毒疾病中所证明的那样。
Chronic wasting disease (CWD) of cervids is a prion disease distinguished by high levels of transmissibility, wherein bodily fluids and excretions are thought to play an important role. Using cervid bioassay and established CWD detection methods, we have previously identified infectious prions in saliva and blood but not urine or feces of CWD+ donors. More recently, we identified very low concentrations of CWD prions in urine of deer by cervid PrP transgenic (Tg[CerPrP]) mouse bioassay and serial protein misfolding cyclic amplification (sPMCA). This finding led us to examine further our initial cervid bioassay experiments using sPMCA. We sought to investigate whether conventional test-negative deer, previously exposed orally to urine and feces from CWD+ sources, may be harboring low level CWD infection not evident in the 19 month observation period. We further attempted to determine the peripheral PrPCWD distribution in these animals. Various neural and lymphoid tissues from conventional test-negative deer were reanalyzed for CWD prions by sPMCA and cervid transgenic mouse bioassay in parallel with appropriate tissue-matched positive and negative controls. PrPCWD was detected in the tissues of orally exposed deer by both sPMCA and Tg[CerPrP] mouse bioassay; each assay revealed very low levels of CWD prions previously undetectable by western blot, ELISA, or IHC. Serial PMCA analysis of individual tissues identified that obex alone was positive in 4 of 5 urine/feces exposed deer. PrPCWD was amplified from both lymphoid and neural tissues of positive control deer but not from identical tissues of negative control deer. Detection of subclinical infection in deer orally exposed to urine and feces (1) suggests that a prolonged subclinical state can exist, necessitating observation periods in excess of two years to detect CWD infection, and (2) illustrates the sensitive and specific application of sPMCA in the diagnosis of low-level prion infection. Based on these results, it is possible that low doses of prions, e.g. following oral exposure to urine and saliva of CWD-infected deer, bypass significant amplification in the LRS, perhaps utilizing a neural conduit between the alimentary tract and CNS, as has been demonstrated in some other prion diseases.
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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
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0.8
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影响因子：
11.8
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影响因子：
11.8
作者：
Angers RC;Seward TS;Napier D;Green M;Hoover E;Spraker T;O'Rourke K;Balachandran A;Telling GC
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Telling GC