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Volume 28, Number 9—September 2022
Research

Rapid Adaptation of Established High-Throughput Molecular Testing Infrastructure for Monkeypox Virus Detection

Dominik Nörz, Hui Ting Tang, Petra Emmerich, Katja Giersch, Nicole Fischer, Stephan Schmiedel, Marylyn M. Addo, Martin Aepfelbacher, Susanne Pfefferle, and Marc LütgehetmannComments to Author 
Author affiliations: University Medical Center Hamburg-Eppendorf, Hamburg, Germany (D. Nörz, H.T. Tang, K. Giersch, N. Fischer, S. Schmiedel, M.M. Addo, M. Aepelbacher, S. Pfefferle, M. Lütgehetmann); Bernhard-Nocht Institute for Tropical Medicine, Hamburg (P. Emmerich, M.M. Addo, S. Pfefferle); German Center for Infection Research, Hamburg (M.M. Addo)

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Table 1

Primer and probe sequences for a dual-target MPXV assay rapidly adapted from established high-throughput SARS-CoV-2 molecular testing infrastructure*

Oligotype Oligo name Sequence, 5′ → 3′ Final concentration, nM
Primers NVAR fwd TCA ACT GAA AAG GCC ATC TAT (2'OMe-G)A 400
NVAR rev GAG TAT AGA GCA CTA TTT CTA AAT CC(2'OMe-C) A 400
MPOX fwd ACG TGT TAA ACA ATG GGT GA(2'OMe-U) G 400
MPOX rev
 AAC ATT TCC ATG AAT CGT AGT (2'OMe-C)C
 400
Probes NVAR P-YAK YakYellow-CCA TGC AAT (BHQ1)ATA CGT ACA AGA TAG TAG CCA AC-BHQ1 75
MPOX P-FAM FAM-TGA ATG AAT (BHQ1)GCG ATA CTG TAT GTG TGG G-BHQ1 75

*Sequences are derived from previously published NVAR assay by Li et al. (9) and MPXV assay by Shchelkunov et al. (10) and adapted for the cobas omni Utility Channel (Roche Diagnostics, https://diagnostics.roche.com). Oligos were custom made by Ella Biotech GmbH (https://www.ellabiotech.com). Indicated final concentration refers to the final oligo concentrations within the reaction mix. 2′O-methyl-RNA bases are indicated as OMe-X. Internal abasic quenchers are indicated as (BHQ1). NVAR, nonvariola orthopoxvirus; MPXV, monkeypox virus.

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References
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