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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
  1. World Health Organization. Disease outbreak news: multi-country monkeypox outbreak in non-endemic countries 29 May 2022 [cited 2022 Jun 7]. https://www.who.int/emergencies/disease-outbreak-news/item/2022-DON388
  2. Antinori  A, Mazzotta  V, Vita  S, Carletti  F, Tacconi  D, Lapini  LE, et al.; INMI Monkeypox Group. Epidemiological, clinical and virological characteristics of four cases of monkeypox support transmission through sexual contact, Italy, May 2022. Euro Surveill. 2022;27:2200421. DOIPubMedGoogle Scholar
  3. Vivancos  R, Anderson  C, Blomquist  P, Balasegaram  S, Bell  A, Bishop  L, et al.; UKHSA Monkeypox Incident Management team; Monkeypox Incident Management Team. Community transmission of monkeypox in the United Kingdom, April to May 2022. Euro Surveill. 2022;27:2200422. DOIPubMedGoogle Scholar
  4. Perez Duque  M, Ribeiro  S, Martins  JV, Casaca  P, Leite  PP, Tavares  M, et al. Ongoing monkeypox virus outbreak, Portugal, 29 April to 23 May 2022. Euro Surveill. 2022;27:2200424. DOIPubMedGoogle Scholar
  5. Bunge  EM, Hoet  B, Chen  L, Lienert  F, Weidenthaler  H, Baer  LR, et al. The changing epidemiology of human monkeypox-A potential threat? A systematic review. PLoS Negl Trop Dis. 2022;16:e0010141. DOIPubMedGoogle Scholar
  6. Reed  KD, Melski  JW, Graham  MB, Regnery  RL, Sotir  MJ, Wegner  MV, et al. The detection of monkeypox in humans in the Western Hemisphere. N Engl J Med. 2004;350:342–50. DOIPubMedGoogle Scholar
  7. Adalja  A, Inglesby  T. A novel international monkeypox outbreak. Ann Intern Med. 2022 May 24 [Epub ahead of print].
  8. Corman  VM, Landt  O, Kaiser  M, Molenkamp  R, Meijer  A, Chu  DK, et al. Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR. Euro Surveill. 2020;25:25. DOIPubMedGoogle Scholar
  9. Li  Y, Olson  VA, Laue  T, Laker  MT, Damon  IK. Detection of monkeypox virus with real-time PCR assays. J Clin Virol. 2006;36:194–203. DOIPubMedGoogle Scholar
  10. Shchelkunov  SN, Shcherbakov  DN, Maksyutov  RA, Gavrilova  EV. Species-specific identification of variola, monkeypox, cowpox, and vaccinia viruses by multiplex real-time PCR assay. J Virol Methods. 2011;175:163–9. DOIPubMedGoogle Scholar
  11. Pfefferle  S, Reucher  S, Nörz  D, Lütgehetmann  M. Evaluation of a quantitative RT-PCR assay for the detection of the emerging coronavirus SARS-CoV-2 using a high throughput system. Euro Surveill. 2020;25:2000152. DOIPubMedGoogle Scholar
  12. Müller  G, Meyer  A, Gras  F, Emmerich  P, Kolakowski  T, Esposito  JJ. Monkeypox virus in liver and spleen of child in Gabon. Lancet. 1988;1:769–70. DOIPubMedGoogle Scholar
  13. Poljak  M, Korva  M, Knap Gašper  N, Fujs Komloš  K, Sagadin  M, Uršič  T, et al. Clinical evaluation of the cobas SARS-CoV-2 test and a diagnostic platform switch during 48 hours in the midst of the COVID-19 pandemic. J Clin Microbiol. 2020;58:e00599–20. DOIPubMedGoogle Scholar
  14. Adler  H, Gould  S, Hine  P, Snell  LB, Wong  W, Houlihan  CF, et al. NHS England High Consequence Infectious Diseases (Airborne) Network. Clinical features and management of human monkeypox: a retrospective observational study in the UK. Lancet Infect Dis. 2022 May 24 [Epub ahead of print].

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