# Unveiling Intra-Clonal Diversity of Monkeypox Virus from Brazil’s First Outbreak Wave

## TL;DR

This study examines the genetic diversity of monkeypox virus within a single skin lesion from Brazil's first outbreak, revealing subtle differences among viral clones.

## Contribution

The study provides new insights into intra-clonal diversity of MPXV from a single patient sample during Brazil's first outbreak wave.

## Key findings

- Three MPXV clones were isolated from a single skin lesion sample, showing distinct plaque characteristics.
- MPXV clones lacked comet-like structures, indicating reduced EEV release compared to VACV-WR.
- Genomic analysis revealed intra-clonal differences but no significant protein-level variations.

## Abstract

The monkeypox virus (MPXV) is an emerging zoonotic pathogen responsible for mpox, a disease characterized by some smallpox-like symptoms, typically mild but occasionally fatal. The largest mpox recorded global outbreak began in May 2022, with over 162,000 cases across 140 countries. Herein, we have analyzed the intra-clonal diversity of MPXV obtained from a single skin lesion sample from a male patient (June 2022). Three viral clones were obtained following phenotypic evaluation of MPXV lysis plaque characteristics over a three-course infection in BSC-40 cells. Unlike the vaccinia virus Western Reserve (VACV-WR) strain, MPXV clones did not produce comet-like structures, suggesting reduced extracellular enveloped virus (EEV) morphotype release, which is associated with viral dissemination. Whole-genome sequencing and assembly identified subtle differences among clones. Comparative genomic analyses, including synteny and single nucleotide variation (SNV) calling, revealed intra-clonal differences and divergence from clade I and II references, although the variety of mutations found did not reveal possible variations at the protein level. Altogether, these findings suggest that although similar, it is possible that distinct MPXV variants may circulate together and can be found in a single exanthematous lesion.