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Pyrosequencing for Microbial Identification and Characterization
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The Identification and Differentiation between Burkholderia mallei and Burkholderia pseudomallei Using One Gene

Damian H Gilling1, Vicki Ann Luna1, Cori Pflugradt1

  • 1Center for Biological Defense, College of Public Health, University of South Florida, Tampa, FL 33612, USA.

International Scholarly Research Notices
|June 29, 2016
PubMed

Insights

Accurate identification of Burkholderia mallei and Burkholderia pseudomallei is crucial. Pyrosequencing assays targeting the fliP gene insertion sequence IS407A provide a sensitive and specific method for differentiating these closely related bacterial species.

Area of Science:

  • Microbiology
  • Genomics
  • Molecular Biology

Background:

  • Burkholderia mallei and Burkholderia pseudomallei are genetically similar, posing identification challenges.
  • Distinguishing between these species and other Burkholderia is critical for diagnostics and research.
  • Previous methods lacked sufficient specificity for reliable differentiation.

Purpose of the Study:

  • To develop a rapid and accurate method for identifying Burkholderia mallei and Burkholderia pseudomallei.
  • To differentiate these species from each other and other related bacteria.
  • To leverage genetic differences in the fliP gene and IS407A insertion sequence.

Main Methods:

  • Utilized pyrosequencing technology to analyze the flagellin P (fliP) gene.
  • Designed specific oligonucleotide primers to target the IS407A insertion site in B. mallei and B. pseudomallei.
  • Tested DNA samples from various Burkholderia species and other bacteria.

Main Results:

  • Pyrosequencing assays demonstrated 100% sensitivity and 100% specificity in identifying B. mallei and B. pseudomallei.
  • The assays reliably detected the presence or absence and orientation of the IS407A insertion within the fliP gene.
  • Successful differentiation was achieved for all tested bacterial samples.

Conclusions:

  • Pyrosequencing assays targeting the IS407A insertion in the fliP gene offer a reliable method for identifying B. mallei and B. pseudomallei.
  • This approach enhances our understanding of the genomic reduction leading to B. mallei.
  • The findings may guide the development of future diagnostic targets.

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