Proof-of-concept for an alternative prion detection method utilizing nanoparticle-enhanced, aptamer-template loop-mediated isothermal amplification

Transmissible spongiform encephalopathies (TSEs), also known as prion diseases, are fatal neurodegenerative disorders found across various host species. Chronic wasting disease (CWD) of cervids is among the TSEs afflicting wildlife populations. Widely distributed in North America, CWD has a devastating impact on cervid populations, and is an economic burden to government agencies and the cervid industry. Because prion diseases have no treatment, surveillance is critical to managing CWD. Current diagnostic approaches infer TSE disease status via measurement of pathogenic prion (PrP^Sc) protein, but lack the sensitivity for detection of CWD prion in antemortem and environmental samples. Next generation prion amplification assays such as Protein Misfolding Cyclic Amplification (PMCA) and Real-Time Quaking-induced Conversion (RT-QuIC) offer sufficient sensitivity for this purpose but suffer from long turnaround times and are resource intensive. As an alternative method, we present a proof-of-concept test that uses Nanoparticle-Enhanced, Aptamer Template Loop-mediated Isothermal Amplification (NEAT-LAMP) using cellular prion (PrP^C) as a model protein. We demonstrate that this fast, sensitive method outperforms current immunoassays with a sensitivity of 10 pM. NEAT-LAMP leverages existing capabilities of diagnostic laboratories and has the potential to improve CWD management efforts.