Development of a polymerase chain reaction assay for the specific identification of Burkholderia mallei and differentiation from Burkholderia pseudomallei and other closely related Burkholderiaceae

Ricky L. Ulrich, Melanie P. Ulrich, Mark A. Schell, H. Stanley Kim, David DeShazer

Research output: Contribution to journalArticlepeer-review

35 Citations (Scopus)

Abstract

Burkholderia mallei and Burkholderia pseudomallei, the etiologic agents responsible for glanders and melioidosis, respectively, are genetically and phenotypically similar and are category B biothreat agents. We used an in silico approach to compare the B. mallei ATCC 23344 and B. pseudomallei K96243 genomes to identify nucleotide sequences unique to B. mallei. Five distinct B. mallei DNA sequences and/or genes were identified and evaluated for polymerase chain reaction (PCR) assay development. Genomic DNAs from a collection of 31 B. mallei and 34 B. pseudomallei isolates, obtained from various geographic, clinical, and environmental sources over a 70-year period, were tested with PCR primers targeted for each of the B. mallei ATCC 23344-specific nucleotide sequences. Of the 5 chromosomal targets analyzed, only PCR primers designed to bimABm were specific for B. mallei. These primers were used to develop a rapid PCR assay for the definitive identification of B. mallei and differentiation from all other bacteria.

Original languageEnglish
Pages (from-to)37-45
Number of pages9
JournalDiagnostic Microbiology and Infectious Disease
Volume55
Issue number1
DOIs
Publication statusPublished - 2006 May
Externally publishedYes

Bibliographical note

Funding Information:
The research described herein was sponsored by NIAID Interagency Agreement Y1-AI-5004-01. The authors thank David Norwood for assistance in performing the real-time PCR analysis on our tissue samples. We would also like to think Katheryn Kenyon and David Heath for reviewing the manuscript.

Keywords

  • Burkholderia
  • Glanders
  • Melioidosis
  • PCR

ASJC Scopus subject areas

  • Microbiology (medical)
  • Infectious Diseases

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