PCR-RFLP analysis of the flagellin sequences for identification of Burkholderia pseudomallei and Burkholderia cepacia from clinical isolates

The flagellin genes of four Burkholderia pseudomallei and two Burkholderia cepacia clinical isolates were studied by a polymerase chain reaction (PCR) -based isolation method using the same pair of primers. The PCR-amplificati on products of the isolates showed a single band of about 1.1 kb, which is similar to a type II B. cepacia flagellin reported previously. In order to distinguish these two species based an the flagellin PCR-amplified products , Pstl and Xhol restriction endonuclease analysis was performed. The result s suggest that there is sufficient diversity within; the flagellin sequence s of the closely related Burkholderia species, B. pseudomallei and B. cepac ia, to enable flagellin-type identification on the basis of the pattern of restriction fragments. In addition, the flagellin gene should be of conside rable use as a genetic marker for clinical identification of these organism s. (C) 1999 Academic Press.