Rapid reconstruction of multidrug-resistant bacterial genomes using CycloneSEQ nanopore sequencing.

Multidrug-resistant (MDR) bacteria commonly harbor highly complex genomes enriched with mobile elements and resistance islands, which are challenging to resolve using short-read sequencing. Here, we evaluated CycloneSEQ, a newly developed Chinese nanopore sequencing platform, for MDR genome reconstruction. Long-read-only assemblies generated near-complete genomes, while hybrid assemblies achieved accuracy exceeding 99.99%. CycloneSEQ effectively resolved complex structures, including tandem repeats within ARG-bearing regions. Notably, updated sequencing chemistry improved single-read accuracy to 96.2% and yielded assembled genomes with 99.98% accuracy. These results demonstrate that CycloneSEQ enables the generation of complete and highly accurate bacterial genomes, highlighting its potential for antimicrobial resistance research and clinical surveillance.