DETECTION OF BURKHOLDERIA-PSEUDOMALLEI IN BLOOD-SAMPLES USING POLYMERASE CHAIN-REACTION

A highly sensitive, specific, rapid and simple method to detect Burkho lderia pseudomallei in blood samples was developed. Two 22-base oligon ucleotide primers, based on sequences from a specific DNA probe, were used for amplification of bacterial DNA by the polymerase chain reacti on (PCR). Amplification with these primers yielded a 178-base pair pro duct in 100 clinical isolates of B. pseudomallei. As little as 0.5 fg of B. pseudomallei DNA was detectable by this method. Experiments invo lving inoculation of the organism into uninfected blood samples showed that the method could he used to detect as few as 1 bacterial cell ml (-1) of whole blood. Non-specific amplification of other bacterial DNA s from 18 samples of bacteria was not observed. Blood samples from sev en patients proven to have melioidosis by haemoculture were positive u sing these primers. The total time required for sample processing, amp lification and visualization was approximately 3.5 h. The high sensiti vity, rapidity and simplicity of this method should make it valuable f or diagnosis, monitoring of drug treatment and for epidemiological stu dies of the melioidosis. (C) 1997 Academic Press Limited.