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

Authors: Christina M. Lazov; Alice Papetti; Graham J. Belsham; Anette Bøtner; Thomas Bruun Rasmussen; Maria Beatrice Boniotti;

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

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.

Country
Denmark
Keywords

Swine enteric coronavirus (SeCoV), Porcine deltacoronavirus (PDCoV), porcine epidemic diarrhea virus (PEDV), Alphacoronavirus, R, swine enteric coronavirus (SeCoV), Recombiant virus, alphacoronavirus, transmissible gastroenteritis virus (TGEV), Article, Virus diagnosis, virus diagnosis; RNA; recombinant virus; alphacoronavirus; deltacoronavirus; porcine epidemic diarrhea virus (PEDV); transmissible gastroenteritis virus (TGEV); swine enteric coronavirus (SeCoV); porcine deltacoronavirus (PDCoV), virus diagnosis, Transmissible gastroenteritis virus (TGEV), Porcine epidemic diarrhea virus (PEDV), RNA, deltacoronavirus, Medicine, porcine deltacoronavirus (PDCoV), /dk/atira/pure/sustainabledevelopmentgoals/good_health_and_well_being; name=SDG 3 - Good Health and Well-being, recombinant virus, Deltacoronavirus

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    popularity
    This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
    Top 10%
    influence
    This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
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    impulse
    This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
    Top 10%
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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
10
Top 10%
Average
Top 10%
Green
gold