Development and Validation of a Panel of One-Step Four-Plex qPCR/RT-qPCR Assays for Simultaneous Detection of SARS-CoV-2 and Other Pathogens Associated with Canine Infectious Respiratory Disease Complex

Canine infectious respiratory disease complex (CIRDC) is the primary cause of in canine population and caused by a wide array viruses bacterial pathogens with coinfections being common. Since its recognition late 2019, Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) has been reported to dogs. Therefore, rapid detection differentiation SARS-CoV-2 from other common viral agents critical public health standpoint. Here, we developed validated panel four one-step multiplex qPCR/RT-qPCR assays for identification twelve associated CIRDC (canine adenovirus-2, distemper virus, herpesvirus-1, influenza A parainfluenza pneumovirus, coronavirus, SARS-CoV-2, Bordetella bronchiseptica, Streptococcus equi subsp. zooepidemicus, Mycoplasma cynos, M. canis), as well three main CIV subtypes (i.e., H3N2, H3N8, H1N1). All demonstrated high specificity analytical sensitivity. This was used test clinical specimens (n = 76) CIRDC-suspected canis, CRCoV were most frequently identified (30.3%, 25.0%, 19.7% samples, respectively). The newly emerging CPnV detected 5.3% samples 30.3%. new comprehensive thus far identifying pathogens, along SARS-CoV-2.