Development of an accessible and scalable qPCR assay for Monkeypox virus detection.

Abigail J PorzucekAbigale M ProctorKatharina E KlinkhammerSarah R TritschMolly A RobertsonJonathan P BashorJack VillaniJorge L SepulvedaChristopher N Mores

Published in: The Journal of infectious diseases (2022)

During the 2022 monkeypox (MPX) outbreak, testing has been limited and results delayed, allowing ongoing transmission. Gold-standard 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 Cq values of 17.46 to 35.59 and titer equivalents 8.01 × 106 to 2.45 × 100 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 labs to increase diagnostic capacity amid the growing MPX outbreak.

Keyphrases

public health
high throughput
loop mediated isothermal amplification
primary care
label free
healthcare
real time pcr
quality improvement
cell cycle
sensitive detection