Abstract
Coronaviruses (CoVs) cause severe respiratory, enteric, and systemic infections in a wide range of hosts, including humans and animals. Porcine epidemic diarrhea virus (PEDV), a member of the Coronaviridae family, is the etiological agent of porcine epidemic diarrhea (PED), a highly contagious intestinal disease affecting pigs of all ages. In this study, we optimized a viability real-time reverse transcriptase polymerase chain reaction (RT-qPCR) for the selective detection of infectious and heat-inactivated PEDV. PEMAX™, EMA™, and PMAxx™ photoactivable dyes along with PtCl4 and CDDP platinum compounds were screened as viability markers using two RT-qPCR assays: firstly, on PEDV purified RNA, and secondly on infectious and thermally inactivated virus suspensions. Furthermore, PMAxx™ pretreatment matched the thermal inactivation pattern obtained by cell culture better than other viability markers. Finally, we further optimized the pretreatment by coupling viability markers with Triton X-100 in inoculated serum resulting in a better estimation of PEDV infectivity than RT-qPCR alone. Our study has provided a rapid analytical tool based on viability RT-qPCR to infer PEDV infectivity with potential application for feed and feed ingredients monitoring in swine industry. This development would allow for greater accuracy in epidemiological surveys and outbreak investigations.
Highlights
Coronaviruses (CoVs) cause severe respiratory, enteric, and systemic infections in a wide range of hosts, including human and animals
Porcine epidemic diarrhea virus (PEDV) was likely introduced into the United States from China in contaminated swine feed ingredients (Scott et al, 2016), and spray-dried plasma proteins (SDPP), an animal by-product used in diets for weaned piglets, was investigated as a potential source of infection after the first-detected porcine epidemic diarrhea (PED) outbreaks in Canada (Pasick et al, 2014)
The indirect transmission is facilitated by the resistance of the virus in the environment (Carvajal et al, 2015), and fecal contaminated feed and feed ingredients have been pointed as being sources of infection in local and transboundary PEDV outbreaks
Summary
Coronaviruses (CoVs) cause severe respiratory, enteric, and systemic infections in a wide range of hosts, including human and animals. PEDV Infectivity by Viability RT-qPCR major concern in swine production after its emergence in North America in 2013, being responsible for worldwide sporadic and large-scale outbreaks (Stevenson et al, 2013; Carvajal et al, 2015; Lee, 2015). Several studies have suggested the importance of contaminated feed and raw feed materials as a potential source of infection in PEDV transmission (Dee et al, 2016; Trudeau et al, 2016; Schumacher et al, 2018). PEDV was likely introduced into the United States from China in contaminated swine feed ingredients (Scott et al, 2016), and spray-dried plasma proteins (SDPP), an animal by-product used in diets for weaned piglets, was investigated as a potential source of infection after the first-detected PED outbreaks in Canada (Pasick et al, 2014). Several studies have confirmed that good manufacturing practices together with at least 2 weeks of storage minimize the risk of infectious PEDV in SDPP (Gerber et al, 2014; Opriessnig et al, 2014; Pujols and Segalés, 2014; Hulst et al, 2019), the detection of PEDV RNA in SDPP still raise concerns in the porcine industry (Foddai et al, 2015)
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