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Multi-locus Variable-number Tandem-repeat Analysis of the Fish-pathogenic Bacterium Yersinia ruckeri by Multiplex PCR and Capillary Electrophoresis
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Allele intersection analysis: a novel tool for multi locus sequence assignment in multiply infected hosts.

Wolfgang Arthofer1, Markus Riegler, Hannes Schuler

  • 1Molecular Ecology Group, Institute of Ecology, University of Innsbruck, Innsbruck, Austria. wolfgang.arthofer@uibk.ac.at

Plos One
|July 27, 2011
PubMed
Summary

This study introduces Allele Intersection Analysis (AIA), a novel method for accurately identifying Wolbachia strains in multiply infected hosts. AIA overcomes limitations of Multi Locus Sequence Typing (MLST) for complex bacterial endosymbiont populations.

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Area of Science:

  • Microbiology
  • Genetics
  • Entomology

Background:

  • Wolbachia are widespread bacterial endosymbionts infecting a large proportion of arthropod species.
  • Accurate identification of Wolbachia strains is crucial for understanding their biology and impact on hosts.
  • Existing Multi Locus Sequence Typing (MLST) methods face challenges in identifying strains in hosts with multiple Wolbachia infections.

Purpose of the Study:

  • To evaluate the limitations of current Multi Locus Sequence Typing (MLST) protocols for identifying Wolbachia strains.
  • To introduce and validate Allele Intersection Analysis (AIA) as a novel method for Wolbachia strain typing.
  • To enable accurate strain assignment in arthropods infected with multiple Wolbachia strains without artificial segregation.

Main Methods:

  • Development and application of Allele Intersection Analysis (AIA) using natural variations in infection patterns.
  • Identification of multiply infected host pairs sharing Wolbachia but differing in only one strain.
  • Utilizing shared MLST sequences within these pairs to assign alleles to distinct Wolbachia strains.

Main Results:

  • Allele Intersection Analysis (AIA) accurately assigns Wolbachia strains in multiply infected hosts.
  • AIA effectively identifies recombination events within Wolbachia populations.
  • The method circumvents the need for artificial strain segregation in host species.

Conclusions:

  • Allele Intersection Analysis (AIA) provides a robust solution for Wolbachia strain typing in complex infections.
  • AIA's principles are adaptable for MLST in other uncultivable bacteria with multiple strain infections.
  • The approach has potential applications in metagenomic sequencing projects for bacterial identification.