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Volume 55, Issue 5

Using real-time PCR to specifically detect Free

Abstract

is the causative agent of human and animal glanders and is a category B biothreat agent. Rapid diagnosis of and immediate prophylactic treatment are essential for patient survival. The majority of current bacteriological and immunological techniques for identifying from clinical samples are time-consuming, and cross-reactivity with closely related organisms (i.e. ) is a problem. In this investigation, two -specific real-time PCR assays targeting the gene ( ntracellular otility A; BMAA0749), which encodes a protein involved in actin polymerization, were developed. The PCR primer and probe sets were tested for specificity against a collection of and isolates obtained from numerous clinical and environmental ( only) sources. The assays were also tested for cross-reactivity using template DNA from 14 closely related species. The relative limit of detection for the assays was found to be 1 pg or 424 genome equivalents. The authors also analysed the applicability of assays to detect within infected BALB/c mouse tissues. Beginning 1 h post aerosol exposure, was successfully identified within the lungs, and starting at 24 h post exposure, in the spleen and liver. Surprisingly, was not detected in the blood of acutely infected animals. This investigation provides two real-time PCR assays for the rapid and specific identification of .

  • Received:
  • Accepted:
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Keyword(s): CT value, crossing threshold value , EPF, end-point fluorescence , MGB probe, minor groove binding probe and p.e., post exposure
SGM
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/content/journal/jmm/10.1099/jmm.0.46350-0
2006-05-01
2024-04-27
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Using real-time PCR to specifically detect Burkholderia mallei
J. Med. Microbiol. 55, 551 (2006); https://doi.org/10.1099/jmm.0.46350-0
/content/journal/jmm/10.1099/jmm.0.46350-0
/content/journal/jmm/10.1099/jmm.0.46350-0
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