修饰蛋白错误折叠环扩增克服实时晃动诱导转换分析物在鹿唾液中检测慢性消耗病朊病毒的抑制剂。
Modified Protein Misfolding Cyclic Amplification Overcomes Real-Time Quaking-Induced Conversion Assay Inhibitors in Deer Saliva To Detect Chronic Wasting Disease Prions.
机构信息
Prion Research Center, Department of Microbiology, Immunology and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado, USA.
Prion Research Center, Department of Microbiology, Immunology and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado, USA
出版信息
J Clin Microbiol. 2018 Aug 27;56(9). doi: 10.1128/JCM.00947-18. Print 2018 Sep.
Chronic wasting disease (CWD), a fatal neurodegenerative prion disease of cervids, has spread across North America and has been detected in The Republic of Korea, Finland, and Norway. CWD appears to spread by horizontal transmission, and prions shed in saliva, feces, and urine are thought to contribute. However, studies investigating the rapid spread of CWD have been hampered by assay inhibitors and a lack of consistent and sensitive means to detect the relatively low levels of prions in these samples. Here we show that saliva frequently contains an inhibitor of the real-time quaking-induced conversion assay (RT-QuIC) and that the inhibitor is a member of the mucin family. To circumvent the inhibitor, we developed a modified protein misfolding cyclic amplification (PMCA) method to amplify CWD prions in saliva that were undetectable or ambiguous by RT-QuIC. Our results reinforce the impact of saliva in horizontal CWD transmission and highlight the importance of detection optimization.
慢性消瘦病（CWD）是一种致命的鹿科动物神经退行性朊病毒病，已在北美传播，并已在韩国、芬兰和挪威检测到。CWD 似乎通过水平传播传播，唾液、粪便和尿液中排出的朊病毒被认为有贡献。然而，研究 CWD 的快速传播受到检测抑制剂的阻碍，并且缺乏一致和敏感的方法来检测这些样本中相对较低水平的朊病毒。在这里，我们表明唾液中经常含有实时震颤诱导转化测定（RT-QuIC）的抑制剂，并且该抑制剂是粘蛋白家族的成员。为了规避抑制剂，我们开发了一种改良的蛋白质错误折叠循环扩增（PMCA）方法，以扩增 RT-QuIC 无法检测或不确定的唾液中的 CWD 朊病毒。我们的结果强调了唾液在水平 CWD 传播中的影响，并强调了检测优化的重要性。
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