Project Summary Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a zoonotic disease that, in addition to its importance in the human pandemic, has caused spillover infection and onward spread in mink, white-tailed deer, and hamsters. New viral variants with different characteristics are constantly emerging, with different characteristics and in some cases, these variants are able to infect species of animals that were not originally susceptible. Therefore, ongoing surveillance through Vet-LIRN is essential for early recognition of spread within animals. Given that companion animals reside in 85 million US households, if SARS-CoV-2 were to gain the ability to spread amongst pets or between pets and humans, there could be immense public health consequence and consequence to wildlife populations. The objective of this proposal is to evaluate and validate novel testing methodologies including validating loop-mediated isothermal amplification (LAMP) in a variety of sample matrices in dogs and cats, and to perform rigorous evaluation of a commercially available ELISA kit. Our specific aims include: Aim 1: Assess the validity of a commercial colorimetric LAMP assay for the qualitative detection of SARS-CoV-2 RNA in 4 novel matrices collected from both dogs and cats. Aim 2: Validate a commercially available ELISA for SARS-CoV-2 antibodies in dogs and cats while building capacity to conduct SARS-CoV-2 serological testing. These aims will provide technician training, equipment, and expertise to provide capacity for SARS-CoV-2 testing in dogs and cats.