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Related Experiment Video

Updated: Jun 1, 2026

Multi-locus Variable-number Tandem-repeat Analysis of the Fish-pathogenic Bacterium Yersinia ruckeri by Multiplex PCR and Capillary Electrophoresis
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Multi-locus Variable-number Tandem-repeat Analysis of the Fish-pathogenic Bacterium Yersinia ruckeri by Multiplex PCR and Capillary Electrophoresis

Published on: June 17, 2019

Multiple-locus variable number tandem repeat analysis for Streptococcus pneumoniae: comparison with PFGE and MLST.

Karin E M Elberse1, Sónia Nunes, Raquel Sá-Leão

  • 1Laboratory for Infectious Diseases and Perinatal Screening, National Institute for Public Health and the Environment (RIVM), Bilthoven, The Netherlands. Karin.Elberse@rivm.nl

Plos One
|June 4, 2011
PubMed
Summary

Multiple-locus variable-number tandem repeat analysis (MLVA) offers a reliable, high-throughput method for tracking pneumococcal strains. This cost-effective technique accurately identifies pneumococcal isolates, aiding disease surveillance efforts.

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Area of Science:

  • Microbiology
  • Genetics
  • Epidemiology

Background:

  • Pneumococcal conjugate vaccines necessitate ongoing surveillance of disease and carriage.
  • Reliable, large-scale genotyping methods are crucial for monitoring population changes.

Purpose of the Study:

  • To introduce and evaluate a high-throughput multiple-locus variable-number tandem repeat analysis (MLVA) for pneumococcal typing.
  • To compare the discriminatory power and congruence of MLVA with pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST).

Main Methods:

  • Developed an 8-BOX loci MLVA method using multiplex PCR and automated DNA sequencing.
  • Applied MLVA to 263 pneumococcal carriage isolates previously characterized by MLST and PFGE.
  • Determined the number of tandem repeats to generate numerical profiles for identification and clustering.

Main Results:

  • MLVA, MLST, and PFGE yielded 164, 120, and 87 types, respectively, with Simpson's diversity indices ≥98.5%.
  • High congruence was observed between MLST and MLVA (Wallace index 0.874 for MLST type, 0.995 for MLST clonal complex).
  • MLVA demonstrated higher discriminatory power than MLST for certain isolates, grouping them by serotype/serogroup.

Conclusions:

  • MLVA is a promising, high-throughput, cost-effective genotyping method for pneumococcal surveillance.
  • MLVA provides a reliable alternative to PFGE and MLST for large-scale applications.
  • The method aids in assessing pneumococcal population structure and changes over time.