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Identification of Rare Bacterial Pathogens by 16S rRNA Gene Sequencing and MALDI-TOF MS
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A possible pitfall in the identification of Burkholderia mallei using molecular identification systems based on the

Lisa D Sprague1, Gregor Zysk, Ralf M Hagen

  • 1Institut für Mikrobiologie der Bundeswehr, Neuherbergstr. 11, 80937 Munich, Germany.

FEMS Immunology and Medical Microbiology
|November 9, 2002
PubMed

Insights

A novel molecular method using the flagellin (fliC) gene was developed to identify Burkholderia mallei and Burkholderia pseudomallei. While effective for species identification, it showed limitations in differentiating between these two closely related bacteria.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Bacterial Pathogenesis

Background:

  • Burkholderia mallei (glanders) and Burkholderia pseudomallei (melioidosis) are closely related, often causing similar clinical syndromes.
  • Accurate identification and differentiation are crucial for effective diagnosis and treatment of these diseases.
  • Existing phenotypic methods can be slow and lack specificity.

Purpose of the Study:

  • To develop a rapid and reliable molecular method for the identification and differentiation of Burkholderia mallei and Burkholderia pseudomallei.
  • To investigate the utility of the flagellin (fliC) gene sequence for distinguishing these species.

Main Methods:

  • Sequencing of the flagellin (fliC) gene in Burkholderia mallei and Burkholderia pseudomallei strains.
  • Design of a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay targeting a specific AvaII restriction site.
  • Testing the assay on B. mallei, B. pseudomallei, and Burkholderia thailandensis strains.

Main Results:

  • A single mutation in the fliC gene was identified that differentiates B. mallei from B. pseudomallei.
  • The PCR-RFLP assay correctly identified all tested B. mallei and B. thailandensis strains.
  • While most B. pseudomallei strains were correctly identified, three strains showed a mutation disrupting the AvaII site, hindering differentiation.

Conclusions:

  • Molecular analysis of the fliC gene sequence allows for reliable identification of B. mallei, B. pseudomallei, and B. thailandensis.
  • The developed PCR-RFLP assay is effective for species identification but has limitations in differentiating B. mallei and B. pseudomallei due to sequence variations.
  • Further refinement of molecular methods is needed for definitive differentiation between these closely related and clinically significant Burkholderia species.

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