Spatiotemporal genomic analysis and risk assessment of the plasmids carrying blaOXA-48-like genes based on a large-scale international dataset

The spread of OXA-48-like carbapenemases represents a major public health challenge. Although previous studies have investigated OXA-48-like carbapenemases risk factors, nosocomial dissemination, and plasmid dynamics, an integrated plasmid-centered framework combining complete plasmid mining, transmission-unit analysis, phylogenetic reconstruction, and machine learning–based risk assessment remains limited. We systematically collected 747 complete plasmid sequences carrying bla OXA-48-like genes from the NCBI database, establishing the largest collections of complete plasmid sequences to date. Using an integrative framework of population genomics, phylogenetic dating, and machine learning, this study aimed to characterize the dissemination patterns, plasmid replicon diversity, transmission units, mobile genetic elements, co-resistance profiles, and risk classification of these plasmid. Plasmids carrying bla OXA-48-like genes were detected across 50 countries on six continents, with bla OXA-48 predominating in Europe, bla OXA-181 in South Asia, and bla OXA-232 largely in Asia. IncL and ColKP3/IncX3 replicons, together with Tn1999. 2 and other MGEs, were central drivers of plasmid maintenance and spread. Sixteen transmission units were defined, with AA068Cluster3 estimated to have originated in the Netherlands around 2005 before expanding to Europe, the Middle East, Asia, and North America. Co-resistance analyses revealed frequent modules involving aminoglycoside and quinolone resistance, with qnrS1 and aph (3’’) -Ib most prevalent. Notably, high-risk transposon structures were often identified in non-clinical environments, underscoring their cross-ecological transmission potential. Machine learning-based classification models showed good internal performance for predefined composite-risk categories, with plasmid mobility, clinical/non-clinical source composition, and host background contributing to the classification results. This study provides a large-scale plasmid-centered genomic analysis of publicly available complete plasmid sequences carrying bla OXA-48-like genes, integrating transmission-unit inference, phylogeographic reconstruction, mobile genetic element and co-resistance profiling, and composite genomic risk stratification. This gene-centered framework may support future One Health–oriented antimicrobial resistance surveillance and prioritization of plasmids with higher dissemination and resistance potential.