Evaluation of RT-LAMP for SARS-CoV-2 Detection in Animal Feces.

Publication date: May 29, 2025

The wide host range, potential lethality, and zoonotic potential of SARS-CoV-2 infection in animals highlights the need for additional surveillance strategies. We validated a commercial, pH-based, colorimetric RT-LAMP assay for the detection of SARS-CoV-2 RNA in animal feces. The comparator assay was 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 min, incubation temperatures of 60 to 70 ^0C, and reaction volumes of 10 to 25 uL. 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%. Blinded method testing organized by an independent laboratory confirmed the satisfactory reproducibility of the assay. To our knowledge, this study represents the first validation of RT-LAMP for SARS-CoV-2 detection in animals. 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, allowing for earlier implementation of control measures.

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Concepts Keywords
Coronaviruses Animals
Lethality animals
Pcr COVID-19
Zoonotic Feces
feces
Humans
Limit of Detection
Molecular Diagnostic Techniques
Reproducibility of Results
RNA, Viral
RNA, Viral
rRT-PCR
RT-LAMP
SARS-CoV-2
SARS-CoV-2
Sensitivity and Specificity
Viral Load

Semantics

Type Source Name
disease IDO assay
pathway REACTOME SARS-CoV-2 Infection
disease MESH SARS-CoV-2 infection
disease IDO host
disease MESH pneumonia
disease MESH infection
disease MESH subclinical infection
disease MESH death
disease MESH interstitial pneumonia
disease MESH acute respiratory distress syndrome
disease MESH fungal infections
drug DRUGBANK Stavudine
disease MESH zoonotic spillover
drug DRUGBANK Gold
disease IDO replication
disease IDO nucleic acid
drug DRUGBANK Phosphate ion
disease IDO pathogen
drug DRUGBANK Water
drug DRUGBANK Cefoxitin
disease MESH gastroenteritis
drug DRUGBANK Sumatriptan
disease MESH viral load
drug DRUGBANK Lauric Acid
drug DRUGBANK Tretamine
disease MESH clinical relevance

Original Article

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