ABSTRACT
Influenza A Virus in swine (IAV-S) is a zoonotic pathogen that is nearly ubiquitous in commercial swine in the USA. Swine possess sialic acid receptors that allow co-infection of human and avian viruses with the potential of pandemic reassortment. We aimed to develop a fast and robust testing method for IAV-S detection on swine farms. Two primers of the RT-LAMP assay were labeled for use in a lateral flow readout. A commercially available lateral flow kit was used to read the amplicon product. With a runtime of â¼ 45â¯minutes, the limit of detection for the assay is comparable with an RT-qPCR Cq less than 35, with a sensitivity of 83.5â¯% and a specificity of 89.6â¯%. This assay allows veterinarians and producers with limited access to diagnostic services to perform and detect Matrix gene amplification on-site with low equipment costs. The time from sample collection to detection is less than one hour, making this method an accessible, convenient, and affordable tool to prevent the spread of zoonotic disease.
ABSTRACT
Point-of-care diagnostic technologies are becoming more widely available for production species. Here, we describe the application of reverse transcription loop-mediated isothermal amplification (RT-LAMP) to detect the matrix (M) gene of influenza A virus in swine (IAV-S). M-specific LAMP primers were designed based on M gene sequences from IAV-S isolated in the USA between 2017 and 2020. The LAMP assay was incubated at 65 °C for 30 min, with the fluorescent signal read every 20 s. The assay's limit of detection (LOD) was 20 M gene copies for direct LAMP of the matrix gene standard, and 100 M gene copies when using spiked extraction kits. The LOD was 1000 M genes when using cell culture samples. Detection in clinical samples showed a sensitivity of 94.3% and a specificity of 94.9%. These results show that the influenza M gene RT-LAMP assay can detect the presence of IAV in research laboratory conditions. With the appropriate fluorescent reader and heat block, the assay could be quickly validated as a low-cost, rapid, IAV-S screening tool for use on farms or in clinical diagnostic labs.
ABSTRACT
This pilot project investigated environmental SARS-CoV-2 presence in seven Midwestern meatpacking plants from May 2020 to January 2021. This study investigated social distancing and infection control practices and incorporated environmental sampling of surfaces and air in employee common areas. All plants increased their social distancing efforts, increased the frequency of cleaning and disinfecting worker areas, and screened for symptomatic people to prevent entry into the workplace. 575 samples from common areas were collected and evaluated with RT-qPCR for the presence of SARS-CoV-2. 42/367 surface samples were positive, while no virus was detected in air samples. Case positive data from the counties surrounding each plant showed peak positive SARS-CoV-2 cases from 12-55 days before the virus was detected in the plant, indicating that environmental sampling is likely a lagging indicator of community and plant infection.
