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Diagnostic microbiology and infectious disease2006; 54(4); 241-247; doi: 10.1016/j.diagmicrobio.2005.09.018

Detection of the reemerging agent Burkholderia mallei in a recent outbreak of glanders in the United Arab Emirates by a newly developed fliP-based polymerase chain reaction assay.

Abstract: A polymerase chain reaction (PCR) assay targeting the flagellin P (fliP)-I S407A genomic region of Burkholderia mallei was developed for the specific detection of this organism in pure cultures and clinical samples from a recent outbreak of equine glanders. Primers deduced from the known fliP-IS407A sequence of B. mallei American Type Culture Collection (ATCC) 23344(T) allowed the specific amplification of a 989-bp fragment from each of the 20 B. mallei strains investigated, whereas other closely related organisms tested negative. The detection limit of the assay was 10 fg for purified DNA of B. mallei ATCC 23344(T). B. mallei DNA was also amplified from various tissues of horses with a generalized B. mallei infection. The developed PCR assay can be used as a simple and rapid tool for the specific and sensitive detection of B. mallei in clinical samples.
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Publication Date: 2006-02-08 PubMed ID: 16466896DOI: 10.1016/j.diagmicrobio.2005.09.018Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research centers around developing a Polymerase Chain Reaction (PCR) test for accurately identifying the Burkholderia mallei bacterium, responsible for glanders in horses, which had shown up during an outbreak in the United Arab Emirates. The newly created PCR assay specifically targets the flagellin P genomic region of the bacterium, ensuring reliable detection in both pure cultures and clinical samples.

Development of the PCR Assay

  • The researchers developed a PCR assay that specifically targets the fliP-IS407A genomic region in Burkholderia mallei. This was made possible by using primers, which are short fragments of DNA and crucial for DNA amplification, derived from the known sequence of the fliP-IS407A of a B. mallei strain held in the American Type Culture Collection (ATCC).
  • This assay was designed to amplify a specific 989-bp fragment from every strain of B. mallei, thus making it specific to this bacterium.
  • Researchers confirmed the specificity of this assay by testing other closely related organisms; these organisms did not show any positive results, therefore ensuring that the test was particular for B. mallei.

Assay Efficiency and Sensitivity

  • The efficiency of the PCR assay was highlighted by its ability to detect as low as 10fg (femtograms) of purified DNA from the B. mallei strain.
  • This sensitivity was further confirmed when the researchers successfully applied the PCR assay to various clinical samples taken from horses afflicted with a generalized B. mallei infection.

Conclusion and Potential Implications

  • Given the success of the PCR assay in the lab and with clinical samples, it presents a quick and direct approach to detecting B. mallei in clinical samples.
  • This could prove invaluable for diagnosing glanders in horses more quickly and accurately and could potentially curtail future outbreaks of the disease.

Cite This Article

APA
Scholz HC, Joseph M, Tomaso H, Al Dahouk S, Witte A, Kinne J, Hagen RM, Wernery R, Wernery U, Neubauer H. (2006). Detection of the reemerging agent Burkholderia mallei in a recent outbreak of glanders in the United Arab Emirates by a newly developed fliP-based polymerase chain reaction assay. Diagn Microbiol Infect Dis, 54(4), 241-247. https://doi.org/10.1016/j.diagmicrobio.2005.09.018

Publication

Diagnostic microbiology and infectious disease
ISSN: 0732-8893
NlmUniqueID: 8305899
Country: United States
Language: English
Volume: 54
Issue: 4
Pages: 241-247

Researcher Affiliations

Scholz, Holger C
  • Bundeswehr Institute of Microbiology, D-80937 Munich, Germany. holger1scholz@bundeswehr.org
Joseph, Marina
    Tomaso, Herbert
      Al Dahouk, Sascha
        Witte, Angela
          Kinne, Joerg
            Hagen, Ralph M
              Wernery, Renate
                Wernery, Ulrich
                  Neubauer, Heinrich

                    MeSH Terms

                    • Animals
                    • Burkholderia mallei / genetics
                    • Burkholderia mallei / isolation & purification
                    • DNA, Bacterial / analysis
                    • Disease Outbreaks / veterinary
                    • Flagellin / genetics
                    • Glanders / epidemiology
                    • Glanders / microbiology
                    • Horses
                    • Polymerase Chain Reaction / methods
                    • United Arab Emirates / epidemiology

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