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Molecular discrimination of Burkholderia pseudomallei and B mallei



Molecular discrimination of Burkholderia pseudomallei and B mallei



Abstracts of the General Meeting of the American Society for Microbiology 102: 261



Burkholderia pseudomallei and B. mallei, respectively are the causative agents of meliodosis and glanders, primarily in animals (both), 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 emerging biological warfare (BW) threat agents. Current primers for detection are based on genes specific to 16S rRNA, rRNA spacer, and LPS. Not much work is reported with respect to identifying unique polymorphic DNA sequences for detection and discriminating B. mallei from B. pseudomallei. Here, we summarize results based on random primer PCR screening approach for development of discriminatory DNA probes for detection of these two pathogens. Two-hundred random primers were screened against these two pathogens and two control DNA from Pseudomonas diminuta and Bacillus cereus. Only 35 out of 200 random primers yielded useful polymorphisms relative to the two pathogens. Nine out of thirty-five primers yielded identical-sized DNA fragments from the two species of Burkholderia. DNA Polymorphisms differentiating between the two pathogens was also identified. Based on these results, it is feasible to develop the genusspecific and species-specific DNA probes.

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