In vitro susceptibility of eighteen clinical isolates of human monkeypox virus to tecovirimat

Abstract

BACKGROUND In 2022, an outbreak of mpox that started in European countries spread worldwide through human-to-human transmission. Cases have been mostly mild, but severe clinical presentations have been reported. In these cases, tecovirimat has been the drug of choice to treat patients with aggravated disease. OBJECTIVES Here we investigated the tecovirimat susceptibility of 18 clinical isolates of monkeypox virus (MPXV) obtained from different regions of Brazil. METHODS Different concentrations of tecovirimat were added to cell monolayers infected with each MPXV isolate. After 72 hours, cells were fixed and stained for plaque visualization, counting, and measurement. The ortholog of F13L gene from each MPXV isolate was polymerase chain reaction (PCR)-amplified, sequenced, and the predicted protein sequences were analyzed. FINDINGS The eighteen MPXV isolates generated plaques of different sizes. Although all isolates were highly sensitive to the drug, two showed different response curves and IC 50 values. However, the target protein of tecovirimat, F13 (VP37), was 100% conserved in all MPXV isolates and therefore does not explain the difference in sensitivity. MAIN CONCLUSIONS Our results support screening different MPXV isolates for tecovirimat susceptibility as an important tool to better use of the restricted number of tecovirimat doses available in low-income countries to treat patients with mpox.

Mpox outbreak in Rio de Janeiro, Brazil: A translational approach

“…Anti‐OPXV IgM detection was not performed. Additionally, MPXV isolation in cell culture was performed from swab samples as previously described 24 . MPXV isolate was fixed in 2.5% glutaraldehyde and analyzed by scanning electron microscopy (SEM) 25 .…”

“…Additionally, MPXV isolation in cell culture was performed from swab samples as previously described. 24 MPXV isolate was fixed in 2.5% glutaraldehyde and analyzed by scanning electron microscopy (SEM). 25 For MPXV genome sequencing, samples were selected (Ct < 28) and PCR amplified using the COVIDSeq kit (Illumina), with modifications as previously described.…”