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-amplification 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 on the flagellin PCR-amplified products, Pst I and Xho I restriction endonuclease analysis was performed. The results suggest that there is sufficient diversity within the flagellin sequences of the closely related Burkholderia species, B. pseudomallei and B. cepacia, to enable flagellin-type identification on the basis of the pattern of restriction fragments. In addition, the flagellin gene should be of considerable use as a genetic marker for clinical identification of these organisms.
Tungpradabkul
S
S
Faculty of Science, Mahidol University, Rama VI Rd., Bangkok, 10400, Thailand. scstp@mucc.mahidol.ac.th
Wajanarogana
S
S
Tunpiboonsak
S
S
Panyim
S
S
D000595:Amino Acid Sequence; D019121:Burkholderia Infections / Q000382:microbiology*; D016956:Burkholderia cepacia / Q000235:genetics* / Q000302:isolation & purification; D016957:Burkholderia pseudomallei / Q000235:genetics* / Q000302:isolation & purification; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D005408:Flagellin / Q000235:genetics*; D005798:Genes, Bacterial; D005819:Genetic Markers; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D013045:Species Specificity
D004269:DNA, Bacterial; D005819:Genetic Markers; D005408:Flagellin