title: Evaluation of molecular typing methods for identification of outbreak-associated Neisseria meningitidis isolates.
abstract: It is essential in an outbreak investigation that strain characterization of Neisseria meningitidis is performed in a rapid and accurate manner. This study evaluated two new molecular typing methods, multiple-locus variable number tandem repeat analysis (MLVA) and repetitive sequence-based PCR (rep-PCR) (DiversiLab; bioMérieux) and compared them with current recommended methodologies. This retrospective study included 36 invasive N. meningitidis serogroup C isolates collected in Sweden 2001 through 2009 and previously subjected to outbreak investigation. All strains were typed with highly variable-MLVA (HV-MLVA) and rep-PCR. The isolates were further characterized by multilocus sequence typing (MLST) and sequencing of the fetA, fHbp, penA, porA and porB genes. The results showed that HV-MLVA had the highest index of diversity (0.99) and rep-PCR had the highest congruence (40%) with the currently recommended typing methods. The HV-MLVA correlated best to the spatiotemporal connections and had the overall highest Adjusted Wallace coefficients, suggesting that HV-MLVA can predict the results of the other typing methods in the study. We therefore suggest that after initial confirmation of species, serogroup and genosubtype, HV-MLVA should be used as the most discriminatory method for first hand investigation of N. meningitidis serogroup C isolates.
authors.affiliation: National Reference Laboratory for Pathogenic Neisseria, Department of Laboratory Medicine, Clinical Microbiology, Örebro University Hospital, Örebro, Sweden. nora-bianka.torosvig@orebroll.se
authors.forename: Bianca
authors.initials: B
authors.lastname: Törös
authors.forename: Sara T
authors.initials: ST
authors.lastname: Hedberg
authors.forename: Susanne
authors.initials: S
authors.lastname: Jacobsson
authors.forename: Hans
authors.initials: H
authors.lastname: Fredlund
authors.forename: Per
authors.initials: P
authors.lastname: Olcén
authors.forename: Paula
authors.initials: P
authors.lastname: Mölling
mesh_terms: D000942:Antigens, Bacterial / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques / Q000379:methods*; D004196:Disease Outbreaks; D005798:Genes, Bacterial; D014644:Genetic Variation; D006801:Humans; D008589:Meningococcal Infections / Q000382:microbiology; D058885:Multilocus Sequence Typing / Q000379:methods; D009345:Neisseria meningitidis / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods; D018272:Porins / Q000235:genetics; D012189:Retrospective Studies; D013548:Sweden / Q000453:epidemiology
chemical_list: D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; C093120:FrpB protein, bacteria; D018272:Porins; C484699:factor H-binding protein, Neisseria meningitidis; C057699:porin protein, Neisseria