Total nucleic acid was extracted from VTM using the Ribo-Virus Viral RNA/DNA extraction kit (Sacace Biotechnologies, Como, Italy) according to the manufacturer’s instructions. Aliquoted nucleic acid and residual samples were stored at -70 C° until further testing with RT-qPCR tests. All samples were tested after one initial freeze-thaw cycle. Commercially available PCR assays were performed according to the manufacturers’ instruction using the Quantstudio 5 Real-Time PCR System (Thermo Fisher Scientific, Waltham, United States)

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