Comparative analysis of two NGS platforms and different databases for analysis of AMR genes

Abstract

The use of antibiotics in human medicine and livestock production has contributed to the widespread occurrence of antimicrobial resistance (AMR). Recognizing the relevance of AMR to human and livestock health, it is important to assess the occurrence of genetic determinants of resistance in medical, veterinary, and public health settings in order to understand risks of transmission and treatment failure. Advances in Next Generation Sequencing (NGS) technologies have had a significant impact on research in microbial genetics and microbiome analyses. Now, strategies for high throughput sequencing from panels of PCR amplicons representing known AMR genes offer opportunities for targeted characterization of complex microbial populations. Aim of the present study was to compare the Illumina MiSeq and Ion Torrent S5 Plus sequencing platforms for use with the Ion AmpliSeq AMR Research Panel in a veterinary/public health setting. All samples were processed in parallel for the two sequencing technologies, subsequently following a common bioinformatics workflow to define the occurrence and abundance of AMR gene sequences. Regardless of sequencing platform, the results were closely comparable with minor differences. The Comprehensive Antibiotic Resistance Database (CARD), QIAGEN Microbial Insight - Antimicrobial Resistance (QMI-AR), Antimicrobial resistance database (AR), and CARD-CLC databases were compared for analysis, with the most genes identified using CARD. Drawing on these results we describe an end-to-end workflow for AMR gene analysis using NGS.