Monkeypox Virus Multiplexed PCR Amplicon Sequencing (PrimalSeq): Enhancing Genomic Surveillance Through Targeted Approaches
Abstract
Background: The global re-emergence of human Monkeypox virus (hMPXV) has emphasized the urgent need for efficient genomic surveillance systems.
Methods: This study evaluates an amplicon-based sequencing approach (PrimalSeq) integrating multiplex PCR with Illumina MiSeq sequencing. Twenty-five clinical samples were analyzed and compared with metagenomic sequencing.
Results: PrimalSeq achieved high genome coverage (>97.8%) and demonstrated superior sensitivity, particularly in samples with high Ct values. It also reduced sequencing costs and turnaround time.
Conclusion: PrimalSeq represents a scalable, cost-effective, and sensitive method for real-time genomic surveillance of hMPXV, particularly in resource-limited settings.