Abstract
BackgroundBurkholderia 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.MethodsSignature sequences for sensitive, specific detection of pathogenic Burkholderia based on published genomes were identified and a qPCR assay was designed and validated.ResultsA 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 genome equivalent per reaction.ConclusionsThe new assay permits rapid detection of pathogenic Burkholderia and combines enhanced sensitivity, species differentiation, and inclusion of an internal control for both DNA extraction and PCR amplification.
Highlights
Burkholderia mallei and B. pseudomallei are two closely related species of highly virulent bacteria that can be difficult to detect
Burkholderia pseudomallei is present in the environment and is a facultative pathogen that causes melioidosis, a glanders-like disease
We have developed a single-reaction quadruplex qPCR assay for rapid, reliable detection of pathogenic Burkholderia
Summary
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, occur elsewhere. Burkholderia pseudomallei is present in the environment and is a facultative pathogen that causes melioidosis, a glanders-like disease. It is a disease of humans and animals in all tropical and sub-tropical regions, but South and Southeast Asia and northern Australia. Cases, included those brought by travelers, occur outside endemic regions [2,3]. Glanders and melioidosis cause diagnostic problems in endemic regions, and even more so when imported into non-endemic areas due to a lack of awareness of these diseases there
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