MALDI-TOF MS: An alternative approach for ribotyping Clostridioides difficile isolates in Brazil.

Area of Science:

• Microbiology
• Infectious Diseases
• Biotechnology

Background:

• Clostridioides difficile infection (CDI) poses a significant healthcare challenge globally.
• Specific C. difficile strains, like BI/NAP1 ribotype 027, have caused widespread outbreaks with high mortality.
• Brazil faces CDI outbreaks from unique ribotypes, yet lacks standard PCR-ribotyping for surveillance.

Purpose of the Study:

• To develop and validate an in-house database using MALDI-TOF MS for identifying C. difficile ribotypes prevalent in Brazilian hospitals.
• To establish a rapid, feasible, and reliable method for epidemiological surveillance of C. difficile in Brazil.

Main Methods:

• Creation of a MALDI-TOF MS database encompassing 19 C. difficile ribotypes (13 global, 6 Brazilian).
• Spectra analysis using BioNumerics software to cluster ribotypes.
• Identification of 13 key biomarkers for ribotype-specific profiling.

Main Results:

• MALDI-TOF MS successfully discriminated all 19 C. difficile ribotypes, forming distinct clusters.
• A panel of 13 biomarkers was identified, with 73% accuracy in ribotype assignment in a double-blind assessment.
• High accuracy (94%-100%) was achieved for the epidemic strain 027 and Brazilian-specific ribotypes.

Conclusions:

• MALDI-TOF MS is a viable alternative for C. difficile epidemiological surveillance in Brazil.
• The developed database and biomarker approach enable rapid identification of local and global C. difficile strains.
• This method supports enhanced public health strategies against CDI outbreaks in Brazil.