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Introduction <p>Diphtheria is a severe, vaccine-preventable infection caused mainly by <i>Corynebacterium diphtheriae</i>, but other species within the <i>C. diphtheriae</i> complex, including <i>C. ulcerans</i>, can also carry the diphtheria toxin (DT) coding gene (<i>tox</i>). Strains classified as “nontoxigenic toxin-gene bearing” (NTTB) harbor the <i>tox</i> gene but do not produce functional DT, although reactivation remains possible. This study reports the first isolation of a NTTB <i>C. ulcerans</i> strain from a diphtheria case in Brazil.</p> Methods <p>A bacterial strain was isolated in Minas Gerais (2019) and subjected to phenotypic, molecular, and phylogenetic analyses. Identification was performed using MALDI-TOF MS, multiplex PCR (mPCR), and sequencing of the 16S rRNA and <i>rpo</i>B genes, followed by phylogenetic inference. Toxigenicity was assessed by mPCR and the modified Elek test. Antimicrobial susceptibility was determined by the disk diffusion method according BrCAST guidelines.</p> Results <p>MALDI-TOF identified the isolate as <i>C. ulcerans</i>. The identification was confirmed by phylogenetic analysis of the <i>rpo</i>B gene. The strain carried the <i>tox</i> gene, but the Elek test did not show the production of DT, classifying the isolate as NTTB. Antimicrobial testing showed susceptibility to most drugs, with the susceptible, increased exposure profile to benzylpenicillin and ciprofloxacin. No resistance was detected.</p> Conclusion <p>Although non-toxigenic, the isolation of a <i>tox</i>-positive <i>C. ulcerans</i> strain highlights the potential risk for re-emergence of diphtheria in Brazil. Continuous genomic surveillance, investigation of zoonotic reservoirs, and maintenance of high vaccination coverage remain crucial to prevent future outbreaks.</p>

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First isolation of nontoxigenic tox gene-bearing Corynebacterium ulcerans from a diphtheria case in Minas Gerais, Brazil

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  • 发表日期：

    2026年3月2日

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  • Amanda Couto Calazans Silva,
  • Lincoln de Oliveira Sant’Anna,
  • Priscila Cristina Nunes Soares,
  • Tayná do Carmo Sant’Anna Cardoso,
  • Juliana Nunes Ramos,
  • Ana Luíza Mattos-Guaraldi,
  • Louisy Sanches dos Santos

摘要

Introduction

Diphtheria is a severe, vaccine-preventable infection caused mainly by Corynebacterium diphtheriae, but other species within the C. diphtheriae complex, including C. ulcerans, can also carry the diphtheria toxin (DT) coding gene (tox). Strains classified as “nontoxigenic toxin-gene bearing” (NTTB) harbor the tox gene but do not produce functional DT, although reactivation remains possible. This study reports the first isolation of a NTTB C. ulcerans strain from a diphtheria case in Brazil.

Methods

A bacterial strain was isolated in Minas Gerais (2019) and subjected to phenotypic, molecular, and phylogenetic analyses. Identification was performed using MALDI-TOF MS, multiplex PCR (mPCR), and sequencing of the 16S rRNA and rpoB genes, followed by phylogenetic inference. Toxigenicity was assessed by mPCR and the modified Elek test. Antimicrobial susceptibility was determined by the disk diffusion method according BrCAST guidelines.

Results

MALDI-TOF identified the isolate as C. ulcerans. The identification was confirmed by phylogenetic analysis of the rpoB gene. The strain carried the tox gene, but the Elek test did not show the production of DT, classifying the isolate as NTTB. Antimicrobial testing showed susceptibility to most drugs, with the susceptible, increased exposure profile to benzylpenicillin and ciprofloxacin. No resistance was detected.

Conclusion

Although non-toxigenic, the isolation of a tox-positive C. ulcerans strain highlights the potential risk for re-emergence of diphtheria in Brazil. Continuous genomic surveillance, investigation of zoonotic reservoirs, and maintenance of high vaccination coverage remain crucial to prevent future outbreaks.