| Summary: | <i>Pseudomonas aeruginosa</i> is a high-priority bacterial agent that causes healthcare-acquired infections (HAIs), which often leads to serious infections and poor prognosis in vulnerable patients. Its increasing resistance to antimicrobials, associated with SPM production, is a case of public health concern. Therefore, this study aims to determine the antimicrobial resistance, virulence, and genotyping features of <i>P. aeruginosa</i> strains producing SPM-1 in the Northern region of Brazil. To determine the presence of virulence and resistance genes, the PCR technique was used. For the susceptibility profile of antimicrobials, the Kirby–Bauer disk diffusion method was performed on Mueller–Hinton agar. The MLST technique was used to define the ST of the isolates. The <i>exoS<sup>+</sup>/exoU<sup>−</sup></i> virulotype was standard for all strains, with the <i>aprA</i>, <i>lasA</i>, <i>toxA</i>, <i>exoS</i>, <i>exoT,</i> and <i>exoY</i> genes as the most prevalent. All the isolates showed an MDR or XDR profile against the six classes of antimicrobials tested. HRC ST277 played a major role in spreading the SPM-1-producing <i>P. aeruginosa</i> strains.
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