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

Christina M. Lazov, Alice Papetti, Graham J. Belsham, Anette Bøtner, Thomas Bruun Rasmussen*, Maria Beatrice Boniotti*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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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, whereas 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.
Original languageEnglish
Article number1040
JournalPathogens
Volume12
Issue number8
Number of pages14
ISSN2076-0817
DOIs
Publication statusPublished - 2023

Keywords

  • Virus diagnosis
  • RNA
  • Recombiant virus
  • Alphacoronavirus
  • Deltacoronavirus
  • Porcine epidemic diarrhea virus (PEDV)
  • Transmissible gastroenteritis virus (TGEV)
  • Swine enteric coronavirus (SeCoV)
  • Porcine deltacoronavirus (PDCoV)

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