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Multiple locus variable number tandem repeat analysis (MLVA) of the pathogenic intestinal spirochaete Brachyspira

Eugene Neo1, Tom La, Nyree Dale Phillips

  • 1School of Veterinary and Life Sciences, Murdoch University, Murdoch, Western Australia 6150, Australia.

Veterinary Microbiology
|February 9, 2013
PubMed

Insights

A new multiple-locus variable number tandem repeat analysis (MLVA) method effectively differentiates strains of Brachyspira pilosicoli, a bacterium causing intestinal issues in various hosts. This tool aids in rapid molecular epidemiology investigations of Brachyspira pilosicoli.

Area of Science:

  • Veterinary Microbiology
  • Molecular Epidemiology
  • Bacterial Genetics

Background:

  • Brachyspira pilosicoli is an anaerobic intestinal spirochaete implicated in diarrhoea and poor growth in multiple host species.
  • Intestinal spirochaetosis, caused by B. pilosicoli, presents a potential zoonotic risk.
  • Accurate molecular tools are needed to understand the epidemiology of B. pilosicoli infections.

Purpose of the Study:

  • To develop a simple and rapid multiple-locus variable number tandem repeat analysis (MLVA) method for investigating the molecular epidemiology of Brachyspira pilosicoli.
  • To assess the discriminatory power of the developed MLVA method for differentiating B. pilosicoli strains.

Main Methods:

  • Genomic analysis of B. pilosicoli strain 95/1000 identified 22 potential tandem repeat loci using Tandem Repeat Finder.
  • Five variable loci were selected and tested on 10 isolates, then applied to a collection of 119 isolates from diverse hosts and locations.
  • VNTR profiles were generated, and a phylogenetic tree was constructed from allelic profiles.

Main Results:

  • A total of 103 unique VNTR profiles were generated from 119 isolates, with a high discriminatory power of 0.976.
  • The phylogenetic analysis revealed significant diversity among B. pilosicoli strains, with no clear clustering based on host species or geographic origin.
  • The MLVA method demonstrated simplicity, ease of use, and effectiveness in differentiating B. pilosicoli strains, with some epidemiologically linked isolates showing identical or highly similar profiles.

Conclusions:

  • The developed MLVA method is a valuable and practical tool for the rapid molecular typing and epidemiological investigation of Brachyspira pilosicoli.
  • This method can effectively distinguish between different strains of B. pilosicoli, aiding in outbreak investigations and understanding transmission dynamics.
  • The findings highlight the genetic diversity of B. pilosicoli and the utility of MLVA in tracking its spread across different hosts and regions.

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