Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Multiplex Real-Time RT-PCR Assays for Detection and Differentiation of Porcine Enteric Coronaviruses

Christina M. Lazov
ORCID logo ,
Alice Papetti
ORCID logo ,
Graham J. Belsham
ORCID logo ,
Anette Bøtner
ORCID logo ,
Thomas Bruun Rasmussen
* ,
Maria Beatrice Boniotti
* ORCID logo
Version 1 : Received: 28 June 2023 / Approved: 29 June 2023 / Online: 29 June 2023 (13:22:32 CEST)

A peer-reviewed article of this Preprint also exists.

Lazov, C.M.; Papetti, A.; Belsham, G.J.; Bøtner, A.; Rasmussen, T.B.; Boniotti, M.B. Multiplex Real-Time RT-PCR Assays for Detection and Differentiation of Porcine Enteric Coronaviruses. Pathogens 2023, 12, 1040. Lazov, C.M.; Papetti, A.; Belsham, G.J.; Bøtner, A.; Rasmussen, T.B.; Boniotti, M.B. Multiplex Real-Time RT-PCR Assays for Detection and Differentiation of Porcine Enteric Coronaviruses. Pathogens 2023, 12, 1040.

Abstract

It is important to be able to detect and differentiate between distinct porcine enteric coronaviruses that can cause similar diseases. However, the existence of naturally occurring recombinant coronaviruses such as swine enteric coronavirus (SeCoV) can give misleading results with currently used diagnostic methods. Therefore, we have developed and validated three duplex real-time quantitative RT-PCR assays for the simultaneous detection of, and differentiation between, porcine epidemic diarrhea virus (PEDV) and SeCoV. Transmissible gastroenteritis virus (TGEV) is also detected by two out of these three assays. In addition, a novel triplex assay was set up that was able to detect and differentiate between these alphacoronaviruses and the porcine deltacoronavirus (PDCoV). The validated assays have low limits of detection, close to 100% efficiency and were able to correctly identify the presence of PEDV and SeCoV in 55 field samples, while 20 samples of other pathogens did not give a positive result. Implementing one or more of these multiplex assays into the routine diagnostic surveillance for PEDV will ensure that the presence of SeCoV, TGEV and PDCoV will not go unnoticed.

Keywords

Virus diagnosis; RNA; recombinant virus; alphacoronavirus; deltacoronavirus; porcine epidemic diarrhea virus (PEDV); transmissible gastroenteritis virus (TGEV); swine enteric coronavirus (SeCoV); porcine deltacoronavirus (PDCoV)

Subject

Biology and Life Sciences, Virology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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