Use of a multilocus variable-number tandem repeat analysis method for molecular subtyping and phylogenetic analysis of Neisseria meningitidis isolates

BackgroundThe multilocus variable-number tandem repeat (VNTR) analysis (MLVA) technique has been developed for fine typing of many bacterial species. The genomic sequences of Neisseria meningitidis strains Z2491, MC58 and FAM18 have been available for searching potential VNTR loci by computer software. In this study, we developed and evaluated a MLVA method for molecular subtyping and phylogenetic analysis of N. meningitidis strains.ResultsA total of 12 VNTR loci were identified for subtyping and phylogenetic analysis of 100 N. meningitidis isolates, which had previously been characterized by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing. The number of alleles ranges from 3 to 40 for the 12 VNTR loci; theoretically, the numbers of alleles can generate more than 5 × 1011 MLVA types. In total, 93 MLVA types were identified in the 100 isolates, indicating that MLVA is powerful in discriminating N. meningitidis strains. In phylogenetic analysis with the minimal spanning tree method, clonal relationships, established with MLVA types, agreed well with those built with ST types.ConclusionOur study indicates that the MLVA method has a higher degree of resolution than PFGE in discriminating N. meningitidis isolates and may be a useful tool for phylogenetic studies of strains evolving over different time scales.

[1]  D. Scheifele,et al.  Meningococcal disease and vaccination in North America , 2001, Journal of paediatrics and child health.

[2]  N. Saunders,et al.  Diversity in coding tandem repeats in related Neisseria spp. , 2003, BMC Microbiology.

[3]  T. Popović,et al.  Evaluation of Pulsed-Field Gel Electrophoresis in Epidemiological Investigations of Meningococcal Disease Outbreaks Caused byNeisseria meningitidis Serogroup C , 2001, Journal of Clinical Microbiology.

[4]  G. Benson,et al.  Tandem repeats finder: a program to analyze DNA sequences. , 1999, Nucleic acids research.

[5]  T. Popović,et al.  Serogroup W-135 Meningococcal Disease during the Hajj, 2000 , 2003, Emerging infectious diseases.

[6]  N. Mouchel,et al.  Experimentally revised repertoire of putative contingency loci in Neisseria meningitidis strain MC58: evidence for a novel mechanism of phase variation , 2003, Molecular microbiology.

[7]  D. Boxrud,et al.  Multilocus Variable-Number Tandem Repeat AnalysisDistinguishes Outbreak and Sporadic Escherichia coli O157:H7Isolates , 2003, Journal of Clinical Microbiology.

[8]  Paul Keim,et al.  Identification and Characterization of Variable-Number Tandem Repeats in the Yersinia pestis Genome , 2001, Journal of Clinical Microbiology.

[9]  L. Price,et al.  Multiple-Locus Variable-Number Tandem Repeat Analysis Reveals Genetic Relationships within Bacillus anthracis , 2000, Journal of bacteriology.

[10]  M. A. Machado,et al.  Differentiation of Strains of Xylella fastidiosa by a Variable Number of Tandem Repeat Analysis , 2001, Applied and Environmental Microbiology.

[11]  A. Devoy,et al.  Stability of PorA during a Meningococcal Disease Epidemic , 2005, Journal of Clinical Microbiology.

[12]  R. G. Hewinson,et al.  Discrimination of Mycobacterium tuberculosis complex bacteria using novel VNTR-PCR targets. , 2002, Microbiology.

[13]  L. Harrison,et al.  Group A meningococcal carriage in travelers returning from Saudi Arabia. , 1988, JAMA.

[14]  F. Denoeud,et al.  Variable Number of Tandem Repeats in Salmonella enterica subsp. enterica for Typing Purposes , 2004, Journal of Clinical Microbiology.

[15]  L. Price,et al.  Erratum: Multiple-locus variable-number tandem repeat analysis reveals genetic relationships within Bacillus anthracis (Journal of Bacteriology (2000) 182:10 (2928-2936)) , 2000 .

[16]  D. Caugant,et al.  Use of Variable-Number Tandem Repeats To Examine Genetic Diversity of Neisseria meningitidis , 2005, Journal of Clinical Microbiology.

[17]  L. Smythe,et al.  Annals of Clinical Microbiology and Antimicrobials Development of a Multiple-locus Variable Number of Tandem Repeat Analysis (mlva) for Leptospira Interrogans and Its Application to Leptospira Interrogans Serovar Australis Isolates from Far North Queensland, Australia , 2022 .

[18]  M. Jennings,et al.  Genetic characterization of pilin glycosylation and phase variation in Neisseria meningitidis , 2003, Molecular microbiology.

[19]  C. Chiou,et al.  Bmc Infectious Diseases Molecular Epidemiology and Emergence of Worldwide Epidemic Clones of Neisseria Meningitidis in Taiwan , 2022 .

[20]  M. Achtman,et al.  Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Paul Keim,et al.  Francisella tularensis Strain Typing Using Multiple-Locus, Variable-Number Tandem Repeat Analysis , 2001, Journal of Clinical Microbiology.

[22]  Lori A. S. Snyder,et al.  Comparative whole-genome analyses reveal over 100 putative phase-variable genes in the pathogenic Neisseria spp. , 2001, Microbiology.

[23]  Paul Vauterin,et al.  Multiple-Locus Variable-Number Tandem Repeat Analysis of Dutch Bordetella pertussis Strains Reveals Rapid Genetic Changes with Clonal Expansion during the Late 1990s , 2004, Journal of bacteriology.

[24]  Janetta Top,et al.  Multiple-Locus Variable-Number Tandem Repeat Analysis, a Novel Typing Scheme To Study the Genetic Relatedness and Epidemiology of Enterococcus faecium Isolates , 2004, Journal of Clinical Microbiology.