Development of a TaqMan Real-Time Polymerase Chain Reaction Assay for Detection of Burkholderia mallei

Burkholderia mallei is the causative agent of glanders, a highly contagious and reemerging zoonotic disease. B. mallei is closely related to Burkholderia pseudomallei, the etiologic agent of melioidosis. The genetic proximity between B. mallei and B. pseudomallei had always been a problem in development of specific molecular detection tests for B. mallei. In the present investigation, we describe a rapid, sensitive, and specific TaqMan real-time polymerase chain reaction (PCR) assay for detection of B. mallei. The TaqMan real-time PCR assay could detect as little as 1 picogram of B. mallei genomic DNA and 100 copies of the target DNA cloned in pGEM-T Easy vector. In spiked human blood, the assay could detect as few as 5.5 × 103 CFU/mL of B. mallei. The assay did not cross-react with other bacterial strains used in the study and was found specific for B. mallei. TaqMan real-time PCR assay can dramatically decrease the turnaround time for diagnostic results, and hence, appropriate action can be initiated at early stage to control and contain the B. mallei infection.