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Multiplex PCR to Differentiate Monkeypox Virus Clades
Christopher T. Williams, Alessandra Romero-Ramirez, Adeleye Adesola Semiu, Samuel Oluwafunmbi Ifabumuyi, Caitlin Greenland-Bews, Susan Gould, Dominic Wooding, Collette Allen, Anushri Somasundaran, Nicodemus Nnabuike Mkpuma, Dorcas Gado, Jolly Amoche Adole, Abdulakeem Eniola Amoo, Abisola Ajoke Adeyemi, Laure Bosquillon de Jarcy, Christine Goffinet, Jake Dunning, Malcolm G. Semple, Esto Bahizire, Afolabi Akinpelu, Thomas E. Fletcher, Ana Cubas-Atienzar, Cristina Leggio, Adeyinka Adedeji, Adesuyi A. Omoare
2, Thomas Edwards
2
, and
The International Severe Acute Respiratory and
Emerging Infection Consortium Investigators,1
Author affiliation: Liverpool School of Tropical Medicine, Liverpool, UK (C.T. Williams, A. Romero-Ramirez, C. Greenland-Bews, S. Gould, D. Wooding, A. Somasundaran, A. Cubas-Atienzar, T. Edwards); Nigeria Centre for Disease Control and Prevention, Abuja, Nigeria (A.A. Semiu, S.O. Ifabumuyi, A. Akinpelu, A.A. Omoare); UK Health Security Agency, Porton Down–Salisbury, UK (C. Allen, C. Leggio); National Veterinary Research Institute, Vom, Nigeria (N.N. Mkpuma, D. Gado, J.A. Adole, A. Adedeji, T. Edwards); Nigeria Centre for Disease Control and Prevention Central Public Health Laboratory, Yaba, Nigeria (A.E. Amoo, A.A. Adeyemi); Charité–Universitätsmedizin Berlin, Berlin, Germany (L. Bosquillon de Jarcy); Berlin Institute of Health, Berlin (L. Bosquillon de Jarcy); Liverpool School of Tropical Medicine, Liverpool, UK (L. Bosquillon de Jarcy, C. Goffinet); University of Oxford, Oxford, UK (J. Dunning); Infectious Diseases Department, Royal Free London NHS Foundation Trust, London, UK (J. Dunning); University of Liverpool, Liverpool (M.G. Semple); Catholic University of Bukavu, Bugabo Campus, Kinshasa, Democratic Republic of the Congo (E. Bahizire); Liverpool School of Tropical Medicine, Liverpool (T.E. Fletcher)
Main Article
Figure
Figure. Standard curves of each monkeypox virus PCR target for multiplex PCR to differentiate monkeypox virus clades. A) Synthetic DNA containing clade I amplicon (R2 0.997, efficiency 98.1%). B) DNA extracted from stock of clade Ia virus (clade I curve: R2 0.998, efficiency 96.4%; non-IIb curve: R2 0.992, efficiency 103.394%). C) DNA extracted from stock of clade Ib virus (clade I curve: R2 0.986, efficiency 98.858%; non-IIb curve: R2 0.996, efficiency 93.9%; Ib curve: R2 0.996, efficiency 95.067%). D) Synthetic DNA containing non-IIb amplicon (R2 0.994, efficiency 97%). E) Synthetic DNA containing IIb amplicon (R2 0.98, efficiency 109.2%). F) DNA extracted from stock of lineage B.1 virus (IIb curve: R2 0.999, efficiency 101.4%; lineage B.1 curve: R2 0.996, efficiency 96.6%). Ct, cycle threshold, R2, coefficient of determination.
Main Article
Page created: January 26, 2026
Page updated: February 09, 2026
Page reviewed: February 09, 2026
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