Mpox virus variant surveillance
Using NextGenPCR™ library preparation for Nanopore sequencing
Rapid genomic surveillance of zoonotic mpox virus (MPXV) infections can provide valuable insights into geographic/demographic viral transmission and guide public health interventions. In current MPXV sequencing approaches, nucleotides isolated from samples are directly sequenced on a nanopore sequencer. The existing methodology is inefficient due to its low genomic coverage and sequencing depth of the fragments of interest and costly because samples cannot be multiplexed.
To aid laboratories with their MPXV genetic surveillance efforts, Molecular Biology Systems has developed an ultrafast library preparation protocol for the analysis of MPXV genomes from samples. Amplification of the 190kb MPXV genome was achieved by using PCR-based amplicon tiling of 2.5kb segments (88 amplicons total) inspired by the SARS-CoV-2 Midnight Protocol and the ARTIC network. With NextGenPCR™ ultrafast PCR chemistry, the complete PCR cycling program of the protocol can be completed in under 42 minutes.
The full laboratory protocol can be viewed here : https://www.protocols.io/
The MPXV primer panel
The composition of the oligonucleotide primer panel was developed by Martin Schou Pedersen from the Department of Clinical Microbiology at Rigshospitalet, Copenhagen University Hospital, Denmark and the NextGenPCR™ MPXV library preparation protocol was tested on 40 samples in close collaboration with Matthijs Welkers and Marcel Jonges from the Dept. of Medical Microbiology & Infection Prevention at Amsterdam UMC (location AMC), The Netherlands.
Using NextGenPCR™ MPXV library preparation up to 20 samples could be multiplexed in a single nanopore flowcell with greatly improved sequencing depth and coverage owing to its PCR tiling based target enrichment. This additionally results in lower costs per sample. The NextGenPCR™ MPXV library preparation method was successfully employed to sequence the first known case of MPXV infection in The Netherlands.
Products:
- NextGenPCR™ MPXV Amplification Primer Panel 96RXN
Cat. 50025-1 - NextGenPCR™ MPXV Sequencing Library Prep Kit 96RXN
Cat. 50027-1
Accelerate your mpox virus sequencing with NextGenPCR™
In the rapidly evolving landscape of infectious diseases, timely and accurate detection is crucial. The emergence of mpox virus (MPXV) has created an urgent need for efficient and reliable genomic surveillance tools. NextGenPCR™ delivers unparalleled speed, allowing you to complete a full 30-cycle PCR runs in just minutes.
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