Pre-Clinical Evaluation of a Real-Time PCR Assay on a Portable Instrument as a Possible Field Diagnostic Tool: Experiences from the Testing of Clinical Samples for African and Classical Swine Fever Viruses.
African swine fever (ASF) and classical swine fever (CSF) are two highly infectious transboundary animal diseases (TADs) that are serious threats to the pig industry worldwide, including in China, the world's largest pork producer. In this study, a duplex real-time PCR assay was developed for the rapid detection and differentiation of African swine fever virus (ASFV) and classical swine fever virus (CSFV). The assay was performed on a portable, battery-powered PCR thermocycler with a low sample throughput (termed as 'T-COR4 assay'). The feasibility and reliability of the T-COR4 assay as a possible field method was investigated by testing clinical samples collected in China. When evaluated with reference materials or samples from experimental infections, the assay performed in a reliable manner, producing results comparable to those obtained from stationary PCR platforms. Of 59 clinical samples, 41 had results identical to a two-step CSFV real-time PCR assay. No ASFV was detected in these samples. The T-COR4 assay was technically easy to perform and produced results within 3 h, including sample preparation. In combination with a simple sample preparation method, the T-COR4 assay provides a new tool for the field diagnosis and differentiation of ASF and CSF, which could be of particular value in remote areas.
National Veterinary Institute (SVA), Uppsala, Sweden.|The OIE Collaborating Centre for the Biotechnology-based Diagnosis of Infectious Diseases in Veterinary Medicine, Uppsala, Sweden.|Joint Laboratory for Veterinary Microbiology between SVA and HVRI, Uppsala, Sweden.
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Liu
Joint Laboratory for Veterinary Microbiology between SVA and HVRI, Uppsala, Sweden.|Division of High-consequence Swine Viral Diseases, Harbin Veterinary Research Institute (HVRI), Chinese Academy of Agricultural Sciences, Harbin, China.
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Luo
Centre de Recerca en Sanitat Animal (CReSA) - Institut de Recerca i Tecnologia Agroalimentàries (IRTA), Barcelona, Spain.|Departament de Sanitat i Anatomia Animals, Facultat de Veterinària (UAB), Barcelona, Spain.
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Accensi
Centre de Recerca en Sanitat Animal (CReSA) - Institut de Recerca i Tecnologia Agroalimentàries (IRTA), Barcelona, Spain.
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Ganges
Centre de Recerca en Sanitat Animal (CReSA) - Institut de Recerca i Tecnologia Agroalimentàries (IRTA), Barcelona, Spain.|Departament de Sanitat i Anatomia Animals, Facultat de Veterinària (UAB), Barcelona, Spain.
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Rodríguez
Qingdao Agricultural University, Qingdao, China.
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Shan
National Veterinary Institute (SVA), Uppsala, Sweden.|The OIE Collaborating Centre for the Biotechnology-based Diagnosis of Infectious Diseases in Veterinary Medicine, Uppsala, Sweden.
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Ståhl
Joint Laboratory for Veterinary Microbiology between SVA and HVRI, Uppsala, Sweden.|Division of High-consequence Swine Viral Diseases, Harbin Veterinary Research Institute (HVRI), Chinese Academy of Agricultural Sciences, Harbin, China.
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Qiu
National Veterinary Institute (SVA), Uppsala, Sweden.|The OIE Collaborating Centre for the Biotechnology-based Diagnosis of Infectious Diseases in Veterinary Medicine, Uppsala, Sweden.|Department of Biomedical Sciences and Veterinary Public Health (BVF), Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.
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Belák
D000357:African Swine Fever / Q000175:diagnosis* / Q000821:virology; D000358:African Swine Fever Virus / Q000235:genetics / Q000302:isolation & purification*; D000818:Animals; D006691:Classical Swine Fever / Q000175:diagnosis* / Q000821:virology; D006692:Classical Swine Fever Virus / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine
African swine fever virus; classical swine fever virus; field test; on-site diagnosis; portable real-time PCR cycler