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

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

doi:10.1093/infdis/jiac414

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

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

Authors

Abigail J Porzucek¹, Abigale M Proctor¹, Katharina E Klinkhammer¹, Sarah R Tritsch¹, Molly A Robertson¹, Jonathan P Bashor¹, Jack Villani², Jorge L Sepulveda², Christopher N Mores¹

Affiliations

¹ Department of Global Health, Milken Institute School of Public Health, George Washington University, Washington, District of Columbia, USA.
² George Washington University Public Health Laboratory, School of Medicine and Health Sciences, George Washington University, Washington, District of Columbia, USA.

PMID: 36221256
DOI: 10.1093/infdis/jiac414

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Diagnostic Tests, Routine* / methods
Humans
Monkeypox virus / genetics
Mpox (monkeypox)* / diagnosis
Polymerase Chain Reaction*
Reproducibility of Results
Sensitivity and Specificity