Application of the Ibis-T5000 pan-Orthopoxvirus assay to quantitatively detect monkeypox viral loads in clinical specimens from macaques experimentally infected with aerosolized monkeypox virus.
Monkeypox virus (MPXV), a member of the family Poxviridae and genus Orthopoxvirus, causes a smallpox-like disease in humans. A previously described pan-Orthopoxvirus assay, based on a broad-range polymerase chain reaction (PCR) coupled with electrospray ionization mass spectrometry (PCR/ESI-MS), was evaluated for its ability to detect MPXV from spiked human and aerosol-infected cynomolgous macaque (Macaca fascicularis) samples. Detection of MPXV DNA from macaque tissue, blood, and spiked human blood by the PCR/ESI-MS pan-Orthopoxvirus assay was comparable, albeit at slightly higher levels, to the current gold standard method of real-time PCR with the pan-Orthopoxvirus assay and had a limit of detection of 200 plaque-forming units. Furthermore, the platform was able to distinguish MPXV and vaccinia viruses that were spiked into macaque blood samples at various concentrations. This platform provides a new tool for the diagnosis and monitoring of orthopoxviral loads during vaccine or antiviral studies, but also could provide rapid identification during natural outbreaks or bioterrorism attacks.
U.S. Army Medical Research Institute of Infectious Diseases, 1301 Ditto Avenue, Fort Detrick, MD 21702, USA. rebecca.j.grant@amedd.army.mil
Rebecca J
RJ
Grant
Carson D
CD
Baldwin
Aysegul
A
Nalca
Scott
S
Zoll
Lawrence B
LB
Blyn
Mark W
MW
Eshoo
Heather
H
Matthews
Rangarajan
R
Sampath
Chris A
CA
Whitehouse
D000336:Aerosols; D000818:Animals; D006801:Humans; D008252:Macaca fascicularis; D045908:Mpox, Monkeypox / Q000097:blood / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D008993:Monkeypox virus; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D019562:Viral Load
D000336:Aerosols