Isogen-LS (Nippon Gene, Toyama, Japan) was used for RNA preparation from clinical specimens (whole blood, pleural fluid, ascites, pericardial effusion and cerebral fluid), and fcwf-4 cells infected with FCoV (79–1146 strain) according to the manufacturer’s protocol. Total RNA was reverse-transcribed using the PrimeScript RT-PCR kit (Perfect Real Time; Takara Bio, Shiga, Japan), as reported previously [22 (link)].
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