title: Ion torrent personal genome machine sequencing for genomic typing of Neisseria meningitidis for rapid determination of multiple layers of typing information.
abstract: Neisseria meningitidis causes invasive meningococcal disease in infants, toddlers, and adolescents worldwide. DNA sequence-based typing, including multilocus sequence typing, analysis of genetic determinants of antibiotic resistance, and sequence typing of vaccine antigens, has become the standard for molecular epidemiology of the organism. However, PCR of multiple targets and consecutive Sanger sequencing provide logistic constraints to reference laboratories. Taking advantage of the recent development of benchtop next-generation sequencers (NGSs) and of BIGSdb, a database accommodating and analyzing genome sequence data, we therefore explored the feasibility and accuracy of Ion Torrent Personal Genome Machine (PGM) sequencing for genomic typing of meningococci. Three strains from a previous meningococcus serogroup B community outbreak were selected to compare conventional typing results with data generated by semiconductor chip-based sequencing. In addition, sequencing of the meningococcal type strain MC58 provided information about the general performance of the technology. The PGM technology generated sequence information for all target genes addressed. The results were 100% concordant with conventional typing results, with no further editing being necessary. In addition, the amount of typing information, i.e., nucleotides and target genes analyzed, could be substantially increased by the combined use of genome sequencing and BIGSdb compared to conventional methods. In the near future, affordable and fast benchtop NGS machines like the PGM might enable reference laboratories to switch to genomic typing on a routine basis. This will reduce workloads and rapidly provide information for laboratory surveillance, outbreak investigation, assessment of vaccine preventability, and antibiotic resistance gene monitoring.
authors.affiliation: Institute for Hygiene and Microbiology, University of Würzburg, Würzburg, Germany. uvogel@hygiene.uni-wuerzburg.de
authors.forename: Ulrich
authors.initials: U
authors.lastname: Vogel
authors.forename: Rafael
authors.initials: R
authors.lastname: Szczepanowski
authors.forename: Heike
authors.initials: H
authors.lastname: Claus
authors.forename: Sebastian
authors.initials: S
authors.lastname: Jünemann
authors.forename: Karola
authors.initials: K
authors.lastname: Prior
authors.forename: Dag
authors.initials: D
authors.lastname: Harmsen
mesh_terms: D000293:Adolescent; D002675:Child, Preschool; D019295:Computational Biology / Q000379:methods; D004196:Disease Outbreaks; D006801:Humans; D007223:Infant; D008589:Meningococcal Infections / Q000453:epidemiology / Q000382:microbiology; D017720:Molecular Epidemiology / Q000379:methods; D058889:Molecular Typing / Q000379:methods*; D038541:Neisseria meningitidis, Serogroup B / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D017422:Sequence Analysis, DNA / Q000379:methods*