Multiplex Real-Time RT-PCR Assays for Detection and Differentiation of Porcine Enteric Coronaviruses
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Multiplex Real-Time RT-PCR Assays for Detection and Differentiation of Porcine Enteric Coronaviruses. / Lazov, Christina M.; Papetti, Alice; Belsham, Graham J.; Bøtner, Anette; Rasmussen, Thomas Bruun; Boniotti, Maria Beatrice.
In: Pathogens, Vol. 12, No. 8, 1040, 2023.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Multiplex Real-Time RT-PCR Assays for Detection and Differentiation of Porcine Enteric Coronaviruses
AU - Lazov, Christina M.
AU - Papetti, Alice
AU - Belsham, Graham J.
AU - Bøtner, Anette
AU - Rasmussen, Thomas Bruun
AU - Boniotti, Maria Beatrice
N1 - Publisher Copyright: © 2023 by the authors.
PY - 2023
Y1 - 2023
N2 - 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.
AB - 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.
KW - alphacoronavirus
KW - deltacoronavirus
KW - porcine deltacoronavirus (PDCoV)
KW - porcine epidemic diarrhea virus (PEDV)
KW - recombinant virus
KW - RNA
KW - swine enteric coronavirus (SeCoV)
KW - transmissible gastroenteritis virus (TGEV)
KW - virus diagnosis
U2 - 10.3390/pathogens12081040
DO - 10.3390/pathogens12081040
M3 - Journal article
C2 - 37624000
AN - SCOPUS:85168998951
VL - 12
JO - Pathogens
JF - Pathogens
SN - 2076-0817
IS - 8
M1 - 1040
ER -
ID: 365819792