Monkeypox Virus Partial-Genome Amplicon Sequencing for Improvement of Genomic Surveillance during Mpox Outbreaks

Mpox is a reemerging infectious disease caused by monkeypox virus (MPXV). Whole-genome sequencing provides comprehensive surveillance of MPXV but is challenging in resource-limited outbreak settings and on clinical samples with low viral load. We developed a low-cost, high-throughput partial-genome sequencing strategy and a freeware Nextflow pipeline for MPXV genomic surveillance. We targeted 2 genomic regions of MPXV by using short overlapping amplicons. This amplicon-based approach generated high-quality sequences over the 2 genomic regions from clinical specimens, including samples with low viral DNA and from formalin-fixed tissues. This partial-genome sequencing approach can determine MPXV subclades and offers an attractive strategy to lower cost and improve MPXV surveillance during outbreaks in mpox-endemic and -nonendemic countries.