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

Summary African swine fever ( ASF ) and classical swine fever ( CSF ) are two highly infectious transboundary animal diseases ( TAD s) 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‐ COR 4 assay’). The feasibility and reliability of the T‐ COR 4 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‐ COR 4 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‐ COR 4 assay provides a new tool for the field diagnosis and differentiation of ASF and CSF , which could be of particular value in remote areas.