Objective    To establish and optimize a method to identify Burkholderia pseudomallei based on primer-specific polymerase chain reaction (PCR).      Methods    The whole genome of Burkholderia pseudomallei international standard strain K96243 was used as a template to amplify the 16S rRNA encoding gene by PCR using specific primers

and the PCR conditions (annealing temperature

annealing time and cycles) were optimized at the same time. This method was then used to detect Burkholderia pseudomallei from other infectious bacteria (Escherichia coli

Shigella dysenteriae

Salmonella paratyphi

Helicobacter pylori

Staphylococcus aureus

Streptococcus group A and Streptococcus group B)

and to identify clinical Burkholderia pseudomallei isolates.      Results    Three pairs of primers were selected to amplify fragments 283-672

760-1 061 and 771-1 193 of 16S rRNA. The optimized PCR conditions were: pre-denaturation at 94 ℃ for 4 min

denaturation at 94 ℃ for 40 s

annealing at 54.5 ℃ for 20 s

and elongation at 72 ℃ for 40 s; there were 24 cycles

with the last elongation time of 1 min. This method was able to rapidly detect Burkholderia pseudomallei from other common clinical infectious bacteria and to identify 8 clinical Burkholderia pseudomallei isolates at a single colony or whole genome level.      Conclusion    The method to identify Burkholderia pseudomallei based on primer-specific PCR is successfully established and optimized

which facilitates efficient diagnosis of Burkholderia pseudomallei and treatment of diseases associated with Burkholderia pseudomallei.