Dynamics of CWD prion detection in feces and blood from naturally infected white-tailed deer.
Dynamics of CWD prion detection in feces and blood from naturally infected white-tailed deer.
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DOI：
10.1038/s41598-023-46929-9
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发表时间：
2023-11-17
影响因子：
4.6
通讯作者：
中科院分区：
文献类型：
--
作者：
慢性消耗性疾病（CWD）是一种影响鹿的朊病毒疾病。目前，CWD的确证性检测是在死后在奥贝和淋巴组织中进行的。大量的证据表明，存在传染性朊病毒的粪便中的慢性消耗病感染的鹿使用体外朊病毒扩增技术和生物测定。在实验条件下，这已经实现了尽快接种后6个月，这表明该样本类型是一个候选人的死前诊断。在本研究中，我们优化了自然感染，临床前白尾鹿粪便样品中的CWD-朊病毒的检测，通过比较旨在浓缩CWD-朊病毒与直接掺入样品的PMCA反应的方案。将该筛选结果与血液中进行的类似分析进行比较。我们的数据表明，CWD-prion检测粪便中使用PMCA是最好的样品预处理的情况下。我们对169份粪便样本进行了筛查，检测CWD-朊病毒的诊断敏感性和特异性分别为54.81%和98.46%。此外，76个粪便样品的PMCA播种活性进行了比较，在匹配的鹿的血液。我们的研究结果表明，粪便和血液中的CWD-朊病毒在临床前晚期增加，当测试PrP 96 GG动物时，在两种样品类型中表现出相似的检测（> 90%灵敏度）。我们的研究结果有助于了解朊病毒分布在不同的生物样品和多态性变异的白尾鹿。这一信息也与目前确定诊断慢性消耗病的平台的努力有关。
Chronic wasting disease (CWD) is a prion disease affecting cervids. Confirmatory testing of CWD is currently performed postmortem in obex and lymphoid tissues. Extensive evidence demonstrates the presence of infectious prions in feces of CWD-infected deer using in vitro prion-amplification techniques and bioassays. In experimental conditions, this has been achieved as soon as 6-month post-inoculation, suggesting this sample type is a candidate for antemortem diagnosis. In the present study, we optimized the detection of CWD-prions in fecal samples from naturally infected, pre-clinical white-tailed deer by comparing protocols aiming to concentrate CWD-prions with direct spiking of the sample into the PMCA reactions. Results of this screening were compared with similar analyses made in blood. Our data shows that CWD-prion detection in feces using PMCA is best in the absence of sample pre-treatments. We performed a screening of 169 fecal samples, detecting CWD-prions with diagnostic sensitivity and specificity of 54.81% and 98.46%, respectively. In addition, the PMCA seeding activity of 76 fecal samples was compared with that on blood of matched deer. Our findings, demonstrate that CWD-prions in feces and blood are increased at late pre-clinical stages, exhibiting similar detection in both sample types (> 90% sensitivity) when PrP96GG animals are tested. Our findings contribute to understand prion distribution across different biological samples and polymorphic variants in white-tailed deer. This information is also relevant for the current efforts to identify platforms to diagnose CWD.
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