Metagenomic next-generation sequencing of rectal swabs for the surveillance of antimicrobial-resistant organisms on the Illumina Miseq and Oxford MinION platforms

Antimicrobial resistance (AMR) is a public health threat where efficient surveillance is needed to prevent outbreaks. Existing methods for detection of gastrointestinal colonization of multidrug-resistant organisms (MDRO) are limited to specific organisms or resistance mechanisms. Metagenomic next-generation sequencing (mNGS) is a more rapid and agnostic diagnostic approach for microbiome and resistome investigations. We determined if mNGS can detect MDRO from rectal swabs in concordance with standard microbiology results. We performed and compared mNGS performance on short-read Illumina MiSeq (N = 10) and long-read Nanopore MinION (N = 5) platforms directly from rectal swabs to detect vancomycin-resistant enterococci (VRE) and carbapenem-resistant Gram-negative organisms (CRO). We detectedEnterococcus faecium(N = 8) andEnterococcus faecalis(N = 2) with associatedvangenes (9/10) in concordance with VRE culture-based results. We studied the microbiome and identified CRO,Pseudomonas aeruginosa(N = 1),Enterobacter cloacae(N = 1), and KPC-producingKlebsiella pneumoniae(N = 1). Nanopore real-time analysis detected thebla(KPC)gene in 2.3 min and provided genetic context (bla(KPC)harbored on pKPC_Kp46 IncF plasmid). Illumina sequencing provided accurate allelic variant determination (i.e.,bla(KPC-2)) and strain typing of theK. pneumoniae(ST-15). We demonstrated an agnostic approach for surveillance of MDRO, examining advantages of both short- and long-read mNGS methods for AMR detection.