2026-09-22T10:43:46Z https://www.ideals.illinois.edu/oai-pmh

oai:www.ideals.illinois.edu:2142/129179 2025-10-20 col_2142_5131 col_2142_16405 com_2142_5130 com_2142_16404 com_2142_14833

application/pdf en text Submission original under an indefinite embargo labeled 'Open Access'. The submission was exported from vireo on 2025-10-19 without embargo terms The student, Aimee Pepper, accepted the attached license on 2025-03-18 at 11:45. The student, Aimee Pepper, submitted this Thesis for approval on 2025-03-18 at 11:54. This Thesis was approved for publication on 2025-03-20 at 10:36. DSpace SAF Submission Ingestion Package generated from Vireo submission #21683 on 2025-10-19 at 18:09:10 Clinical validation of SARS-CoV-2 RT-LAMP in animals Pepper, Aimee W. 2025-03-20 Wang, Leyi Samuelson, Jonathan P Sander, William E SARS-CoV-2 RT-LAMP rRT-PCR feces animals eng The wide host range, pathogenicity, and zoonotic potential of SARS-CoV-2 infection in animals highlights the need for additional surveillance strategies. Shedding of SARS-CoV-2 RNA within the intestinal tract is prolonged during animal infection, suggesting that surveillance could be accomplished non-invasively through nucleic acid amplification of animal feces. We validated a commercial, pH-based, colorimetric, RT-LAMP assay for the detection of SARS-CoV-2 RNA in animal feces, with comparison to the gold standard assay, rRT-PCR. The limit of detection of the RT-LAMP assay was 72 genome copies per reaction. RT-LAMP was highly specific for SARS-CoV-2 and did not detect other human or animal coronaviruses. RT-LAMP was robust, with valid results generated for incubation lengths of 30 to 45 minutes, incubation temperatures of 60 to 70°C, and reaction volumes of 10 to 25 µL. The diagnostic sensitivity was 100% for clinical fecal samples with high viral loads (Ct ≤25), 97.4% for samples with moderate-to-high viral loads (Ct ≤33), and 62% overall (Ct ≤40). The diagnostic specificity was 97.9% overall. Blinded method testing organized by an independent laboratory confirmed the reproducibility of the assay. SARS-CoV-2 RNA could still be detected by RT-LAMP following storage of fecal suspensions with moderate-to-high or high viral loads at -80°C, -20°C, 4°C, or room temperature for up to 28 days. To our knowledge, this study represents the first clinical evaluation of RT-LAMP for SARS-CoV-2 RNA detection in animal samples. RT-LAMP testing could detect SARS-CoV-2 infection more rapidly and at the point-of-care in animals with moderate-to-high viral loads that are likely to be infectious, allowing for earlier implementation of quarantine and control measures to limit viral spread and therapeutic interventions to reduce animal mortality. 2025-05 Thesis https://hdl.handle.net/2142/129179 Copyright 2025 Aimee Pepper Vet Clinical Medicine VMS-Veterinary Clinical Medcne University of Illinois Urbana-Champaign M.S. Thesis