Burkholderia mallei and B. pseudomallei are two closely related
species of highly virulent bacteria that can be difficult to
detect. Pathogenic Burkholderia are endemic in many regions
worldwide and cases of infection, sometimes brought by travelers
from unsuspected regions, also occur elsewhere. Rapid, sensitive
methods for identification of B. mallei and B. pseudomallei are
urgently needed in the interests of patient treatment and
epidemiological surveillance. Signature sequences for sensitive,
specific detection of pathogenic Burkholderia based on published
genomes were identified and a qPCR assay was designed and
validated. A single-reaction quadruplex qPCR assay for the
detection of pathogenic Burkholderia, which includes a marker
for internal control of DNA extraction and amplification, was
developed. The assay permits differentiation of B. mallei and B.
pseudomallei strains, and probit analysis showed a very low
detection limit. Use of a multicopy signature sequence permits
detection of less than 1 gen