Application of qPCR assays for diagnosing causes of viral mink diarrhea. Preliminary results
Abstract
Gastrointestinal (GI) disorders is the main cause for submitting mink (Neovison vison) carcasses for post-mortem examination at the National Veterinary Institute in Denmark and has been described as the predominant cause of disease and mortality in the Danish mink production (Rattenborg et al. 1999). Diarrhea in mink can be caused by infectious agents (virus, bacteria and parasites) and food-related/multifactorial conditions. Known enteric viral infections are mink enteritis virus (MEV) and mink astrovirus. Coronaviruses and caliciviruses have also been implicated as potential causes or contributors to diarrhea in mink. Rotavirus is poorly described in mink, but has previously been demonstrated in feces from mink pups with and without clinical signs (Jorgensen et al. 1996). The pathogenicity of these viruses could be related to viral load, virulence and the age of the mink. Therefore, there is a need for a quantitative diagnostic approach. We have developed new or adapted previously published real-time PCR/RT-PCR assays for MEV, astrovirus, rota- and coronavirus diagnostics. The technical test validation was initially carried out on archived diarrhea samples from diagnosed positive animals and on normal and diarrhea samples from a case-control study. In order to further validate the applicability of the assays, a testing scheme for normal and affected farms was set up and initiated in June 2015. This protocol will allow optimization of test characteristics (sensitivity, specificity and predictive value) and assessment of the validity of using pooled samples in order to reduce test costs.