Real-Time PCR (RT-PCR) Assays for Burkholderia mallei and B. pseudomallei

摘要

Burkholderia pseudomallei and B. mallei, respectively are the causative agents of meliodosis and glanders, primarily in animals (both pathogens), and in humans (commonly the former but sporadically the latter as well). The two pathogens are gram-negative, facultative anaerobe, motile bacilli. There is no known vaccine, and the treatment with antimicrobials is protracted because of natural resistance of these pathogens to commonly used antibiotics. The virulence factors are only now beginning to be elucidated and understood (Rockseidler et al., 2001, Infection and Immunity, 69: 34-44). These pathogens, consequently, are viewed as a serious public health concern. Early detection of these threat pathogens is critical to minimize the potential risk. There is currently no real-time polymerase chain reaction (PCR) assay for detection of both of these pathogens. Primers and probes corresponding to specific genomic regions were designed and tested in an attempt to develop RT- PCR assays for detection of both these pathogens. Here, we report successful development of highly specific RT-PCR assays for detection of both B. mallei and B. pseudomallei cells. To the best of our knowledge, this work constitutes the first report of RT-PCR assays for detection of these pathogens.