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.
Grant
Rebecca J
RJ
U.S. Army Medical Research Institute of Infectious Diseases, 1301 Ditto Avenue, Fort Detrick, MD 21702, USA. rebecca.j.grant@amedd.army.mil
Baldwin
Carson D
CD
Nalca
Aysegul
A
Zoll
Scott
S
Blyn
Lawrence B
LB
Eshoo
Mark W
MW
Matthews
Heather
H
Sampath
Rangarajan
R
Whitehouse
Chris A
CA
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