Optimization of RT-QuIC Assay Duration for Screening Chronic Wasting Disease in White-Tailed Deer

Yilmaz, Gokhan; Morrill, Tamara; Pilot, William; Ward, Cian; Mitchell, Gordon; Soutyrine, Andrei; Dan, Hanhong; Lin, Min; Guan, Jiewen

doi:10.3390/vetsci11020060

Cited by 2 publications

(1 citation statement)

References 28 publications

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“…The monitoring of fluorescence increase was thus related to a positive amplification reaction of prions. The analysis of the positive and negative samples was hence easy and fast compared to amplification using other recombinant proteins [19,54,57].…”

Section: Discussionmentioning

confidence: 99%

Human PrP E219K as a new and promising substrate for RT-QuIC amplification of human prion strains: a first step towards strain discrimination

Marin-Moreno,

Reine,

Jaffrézic

et al. 2024

Preprint

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SummaryPrion diseases are fatal neurodegenerative diseases that affect mammals through the transconformation of a host protein, the prion protein (PrP), into a toxic and pathogenic conformer termed PrPSc. Until now, the diagnosis is only confirmed with a post-mortem histology study of the central nervous system. Among the methods to detect the etiological agent,in vitroamplification techniques have emerged as very sensitive, highly specific and rapid tools, even though some prion strains remain refractory or difficult to amplify. Here we report the use of a new recombinant substrate for Real-Time Quaking Induced Conversion (RT-QuIC), a natural polymorphism of human prion protein with a lysine at position 219 instead of a glutamic acid, PrP E219K. This substrate amplifies the six sporadic human strains responsible for Creutzfeldt-Jakob Disease (CJD) and the strain responsible for its variant form in a few hours and over a large dilution range of the seeds. Moreover, based on the lag time of the amplification reactions, the PrP E219K substrate allows to discriminate between sporadic and variant CJD strains, a first step towards an ante-mortem typing of the prion strain affecting a patient.

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Section: Discussionmentioning

confidence: 99%

Human PrP E219K as a new and promising substrate for RT-QuIC amplification of human prion strains: a first step towards strain discrimination

Marin-Moreno,

Reine,

Jaffrézic

et al. 2024

Preprint

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Evaluating the Diagnostic Efficacy of Using Pooled Samples for Chronic Wasting Disease Testing and Surveillance

Hepker,

Zhang,

Anantharam

et al. 2024

Pathogens

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Disease monitoring informs the opportunities for intervention by natural resource agencies tasked with managing chronic wasting disease (CWD) in wild cervids. However, allocating funds toward testing can reduce those available for education, outreach, and disease reduction. Implementation of more efficient testing strategies can help meet both an expanding need by resource managers and a burgeoning demand from the hunting public in North America. Here, we evaluated the efficacy of pooled testing using the enzyme-linked immunosorbent assay (ELISA), the current screening test used by veterinary diagnostic laboratories in the United States, and real-time quaking-induced conversion (RT-QuIC), an amplification assay that is being evaluated by the U.S. Department of Agriculture but is not yet approved or commercially available. The samples used in this study consisted of medial retropharyngeal lymph nodes (RPLNs) routinely collected by the Iowa Department of Natural Resources during the 2019–2020 surveillance season. The test pools contained tissue from one positive deer diluted in tissue from an increasing number of undetected deer, with each individual contributing an equal tissue volume. ELISA remained positive with pooling thresholds of 1:1, 1:2, 1:4, and 1:9 at a standard volume of tissue homogenate, whereas RT-QuIC remained positive with pooling thresholds of 1:1, 1:2, 1:4, 1:9, 1:19, and 1:49 at a 0.02% tissue dilution. Our results suggest that pooled testing can reduce diagnostic costs multi-fold, and RT-QuIC can be a viable screening test compatible with current field collection standards.

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Optimization of RT-QuIC Assay Duration for Screening Chronic Wasting Disease in White-Tailed Deer

Cited by 2 publications

References 28 publications

Human PrP E219K as a new and promising substrate for RT-QuIC amplification of human prion strains: a first step towards strain discrimination

Human PrP E219K as a new and promising substrate for RT-QuIC amplification of human prion strains: a first step towards strain discrimination

Evaluating the Diagnostic Efficacy of Using Pooled Samples for Chronic Wasting Disease Testing and Surveillance

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