Supplementary Material 5

The use of Multilocus Sequence Typing (MLST) in the identification of Escherichia coli is crucial for understanding its genetic diversity, evolutionary relationships, and epidemiological significance. MLST provides a standardized and highly reproducible method for strain classification by analyzing variations in housekeeping genes, enabling precise identification of distinct sequence types (STs). This approach is particularly important in differentiating pathogenic E. coli strains, such as extraintestinal pathogenic E. coli (ExPEC) and Shiga toxin-producing E. coli (STEC), from commensal isolates. Additionally, MLST helps track the spread of antimicrobial-resistant clones, monitor outbreak sources, and study global strain distribution through publicly available databases like PubMLST. By offering a high-resolution and scalable approach to bacterial typing, MLST plays a vital role in clinical diagnostics, infection control, and evolutionary studies of E. coli.