Amplicon based Genome Sequencing of a Human Monkeypox Virus Isolate from Central Europe During the 2022 Outbreak

We report the genome sequence reads of a human monkeypox virus (hMPXV). The MPXV_NRL 4279/2022 isolate derived from skin lesions of the index case in Prague, Czech Republic. Propagation was conducted under BSL-4 conditions at the National Laboratory of Virology, University of Pecs, Hungary. The virus was passaged on CV-1 cells. DNA extraction was carried out using the Direct-zol DNA/RNA kit (Zymo).A set of 22 multiplex PCR primer pairs was designed to generate app.10 kb overlapping amplicons covering the viral genome.Sequencing libraries were prepared with the SQK-LSK110 kit (Oxford Nanopore Technologies), and barcoding was done using the Barcoding Expansion kit (EXP-NBD196, ONT). The library was loaded onto an R9.4.1 MinION flow cell (FLO-MIN106) and sequenced on a MinION Mk1B device.