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This research was funded by the Spanish Ministry of Science and Innovation grant PID2021-125599OB-100. LC was supported by the Juan de la Cierva Program (2022), the Spanish Ministry of Science and Innovation. AMA was supported by the ICA grant 001-2022. IRTA is supported by the CERCA program/Generalitat de Catalunya.

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Simultaneous Detection of Classical and African Swine Fever Viruses by Duplex Taqman Real-Time PCR Assay in Pigs Infected with Both Diseases

Publicat a:Pathogens. 14 (5): 473- - 2025-05-13 14(5), DOI: 10.3390/pathogens14050473

Autors: Coronado, Liani; Muñoz-Aguilera, Adriana; Wang, Miaomiao; Muñoz, Ivan; Riquelme, Cristina; Heredia, Saray; Stepniewska, Katarzyna; Gallardo, Carmina; Ganges, Llilianne

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The increasing spread of African swine fever (ASF) in recent years and the presence of classical swine fever (CSF) subclinical forms in endemic countries suggests that the possibility of coinfection with ASF virus (ASFV) and CSF virus (CSFV) in pigs cannot be ruled out in areas where both diseases are prevalent. Thus, rapid and reliable diagnosis through molecular testing is essential for the timely implementation of control measures to prevent the spread of these devastating swine diseases. Here, we have coupled two of the most validated PCR assays for the detection of CSFV and ASFV in a single reaction tube. The combination of the two tests for the detection of two target nucleic acids did not affect the analytical sensitivity, and the duplex RT-qPCR assay was comparable with the standard molecular techniques. The detection limits for CSFV RNA and ASFV DNA were 0.12 TCID50/reaction and 0.25 TCID50/reaction, respectively. The test showed high repeatability and reproducibility, the coefficient of variation was below 2%, and excellent performance was demonstrated in clinical samples. The duplex assay shows great potential to become a robust diagnostic tool for the rapid and reliable detection and differentiation of CSFV and ASFV in areas where both viruses may be circulating.

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