Modified Protein Misfolding Cyclic Amplification Overcomes Real-Time Quaking-Induced Conversion Assay Inhibitors in Deer Saliva To Detect Chronic Wasting Disease Prions
| Autor: | Candace K. Mathiason, Nathaniel D. Denkers, Edward A. Hoover, Kristen A. Davenport, Clare E. Hoover |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Microbiology (medical) Protein Folding Saliva Prions animal diseases Biology Sensitivity and Specificity Clinical Veterinary Microbiology Sonication 03 medical and health sciences medicine Animals Feces Transmission (medicine) Deer Mucin Mucins Temperature Chronic wasting disease medicine.disease Virology 030104 developmental biology Wasting Disease Chronic Protein Misfolding Cyclic Amplification Biological Assay Horizontal transmission |
| Zdroj: | Journal of Clinical Microbiology. 56 |
| ISSN: | 1098-660X 0095-1137 |
| DOI: | 10.1128/jcm.00947-18 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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