Tandem repeat markers as novel diagnostic tools for high resolution fingerprinting of Wolbachia

Markus Riegler, Iñaki Iturbe-Ormaetxe, Megan Woolfit, Wolfgang J. Miller, Scott L. O'Neill

    Research output: Contribution to journalArticle

    39 Citations (Scopus)

    Abstract

    Background: Strains of the endosymbiotic bacterium Wolbachia pipientis are extremely diverse both genotypically and in terms of their induced phenotypes in invertebrate hosts. Despite extensive molecular characterisation of Wolbachia diversity, little is known about the actual genomic diversity within or between closely related strains that group tightly on the basis of existing gene marker systems, including Multiple Locus Sequence Typing (MLST). There is an urgent need for higher resolution fingerprinting markers of Wolbachia for studies of population genetics, horizontal transmission and experimental evolution. Results: The genome of the wMel Wolbachia strain that infects Drosophila melanogaster contains inter- and intragenic tandem repeats that may evolve through expansion or contraction. We identified hypervariable regions in wMel, including intergenic Variable Number Tandem Repeats (VNTRs), and genes encoding ankyrin (ANK) repeat domains. We amplified these markers from 14 related Wolbachia strains belonging to supergroup A and were successful in differentiating size polymorphic alleles. Because of their tandemly repeated structure and length polymorphism, the markers can be used in a PCR-diagnostic multilocus typing approach, analogous to the Multiple Locus VNTR Analysis (MLVA) established for many other bacteria and organisms. The isolated markers are highly specific for supergroup A and not informative for other supergroups. However, in silico analysis of completed genomes from other supergroups revealed the presence of tandem repeats that are variable and could therefore be useful for typing target strains. Conclusions: Wolbachia genomes contain inter- and intragenic tandem repeats that evolve through expansion or contraction. A selection of polymorphic tandem repeats is a novel and useful PCR diagnostic extension to the existing MLST typing system of Wolbachia, as it allows rapid and inexpensive high-throughput fingerprinting of closely related strains for which polymorphic markers were previously lacking.
    Original languageEnglish
    Number of pages15
    JournalBMC Microbiology
    DOIs
    Publication statusPublished - 2012

    Open Access - Access Right Statement

    © 2012 Riegler et al; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

    Keywords

    • fingerprints
    • genetics
    • hosts (biology)
    • microbiology
    • phenotype
    • wolbachia

    Fingerprint

    Dive into the research topics of 'Tandem repeat markers as novel diagnostic tools for high resolution fingerprinting of Wolbachia'. Together they form a unique fingerprint.

    Cite this