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. 2023 Apr 26;227(9):1084-1087.
doi: 10.1093/infdis/jiac414.

Development of an Accessible and Scalable Quantitative Polymerase Chain Reaction Assay for Monkeypox Virus Detection

Abigail J Porzucek  1 , Abigale M Proctor  1 , Katharina E Klinkhammer  1 , Sarah R Tritsch  1 , Molly A Robertson  1 , Jonathan P Bashor  1 , Jack Villani  2 , Jorge L Sepulveda  2 , Christopher N Mores  1
Affiliations

Development of an Accessible and Scalable Quantitative Polymerase Chain Reaction Assay for Monkeypox Virus Detection

Abigail J Porzucek et al. J Infect Dis. .

Abstract

During the 2022 monkeypox (MPX) outbreak, testing has been limited and results delayed, allowing ongoing transmission. Gold-standard quantitative polymerase chain reaction (qPCR) diagnostics are difficult to obtain. This research adapted the June 2022 CDC MPX qPCR assay for broad implementation. Validated using MPX stocks in a matrix with multiple sample types, MPX was detected with cycle threshold (Ct) values 17.46-35.59 and titer equivalents 8.01 × 106 to 2.45 × 100 plaque-forming unit (PFU)/mL. The detection limit was 3.59 PFU/mL. Sensitivity and specificity were both 100%. This qPCR assay can be quickly and broadly implemented in research and public health laboratories to increase diagnostic capacity amid the growing MPX outbreak.

Keywords: PCR; diagnostics; emerging infectious disease; global health; monkeypox; outbreak response; qPCR assay.

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Conflict of interest statement

Potential conflicts of interest. C. N. M. has received funding from the Centers for Disease Control and Prevention, World Health Organization, and the German Government; consulting fees from Curative Inc and Singletto Inc; and has stock options with Singletto Inc. All other authors report no potential conflicts. All authors have submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest. Conflicts that the editors consider relevant to the content of the manuscript have been disclosed.

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