Burkholderia pseudomallei are Gram negative highly pathogenic bacteria of humans and
animals causing a multisystemic disease called melioidosis. They have recently gained a lot of interest
from the research community and public health organisations because of their great potential to be
used as an agent of bioterrorism. This has made the search for simple, rapid, accurate and the most
definitive means of their detection, identification and discrimination very critical and necessary. This
article aimed to review the molecular techniques used for detection, identification and differentiation
of B. pseudomallei. Although, culture and isolation techniques maintained their usefulness in
confirming cases of melioidosis, their time limitation (can take up to a week for confirming diagnosis)
leads to the search for rapid and simple techniques. Consequently, serology-based tests have been
developed which are both faster and less sophisticated. However, the presence of high background
titre levels and cross-reaction with other organisms make it less reliable. Thus, efforts have been
directed to explore rapid and accurate molecular techniques and resulting in the development and
validation of various PCR-based identification techniques targeting either single or multiple genes.
Although requiring some level of instrumentation and expertise, PCR-based techniques have been
reported to be very useful in diagnosis of melioidosis. We recommend the 16S rRNA PCR (especially
augmented with other molecular methods such as gene sequencing and analysis) and MLST
techniques for timely detection, identification and differentiation of B. pseudomallei for routine
diagnosis and epidemiological studies respectively.