High-Resolution Molecular Typing of Vancomycin-Resistant Enterococcus faecium from Romania and Bavaria: Combining Enhanced DNA Microarray and Next Generation Sequencing

in: Biotechniques (2026)
Osadare, Ikubun Elizabeth; Ehricht, Ralf; Abdilahi, Abdinasir; Müller, Elke; Lohde, Mara; Diezel, Celia; Collatz, Maximilian; Braun, Sascha D.; Kieninger, Bärbel; Eichner, Anja; Schneider-Brachert, Wulf; Wellhöfer, Thomas; Frankenfeld, Katrin; Dorneanu, Olivia S.; Monecke, Stefan
Antimicrobial resistance poses a significant challenge for infection control, requiring thedevelopment of accurate and high-throughput diagnostic techniques. We expandedand optimized an existing DNA microarray platform for the molecular characterizationof vancomycin-resistant Enterococcus (VRe) by incorporating resistance, virulence,species-specific, and typing markers. the enhanced microarray allows for thesimultaneous analysis of up to 96 strains, providing detailed genetic profiles of clinicalisolates. VRe strains from Romania and Bavaria, Germany, were analyzed, and the resultswere compared to those obtained using traditional typing methods, such as multilocussequence typing (MLSt). Next-generation sequencing (NGS) was used in parallel tovalidate the microarray findings and explore genomic relationships. the microarrayrevealed considerable genetic diversity and potential epidemiological linkages amongisolates. A novel hexadecimal-based nomenclature system was introduced forstandardized and scalable strain classification. Comparative analysis demonstrated thatthe array profiles provided greater discriminatory power and practical resolution thanMLSt. Receiver operating characteristic (RoC) curve analysis of 187 target genes in 220isolates gave diagnostic sensitivity and specificity of 100%. this integrated approachoffers a cost-effective, rapid, and adaptable global VRe surveillance and infection controltool. It provides a practical alternative to conventional typing systems and facilitatesearly detection of outbreaks and emerging clones.

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