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Journal of Clinical Microbiology, June 1999, p. 1906-1912, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Phylogenetic Analysis of Ara+ and Araminus Burkholderia pseudomallei Isolates and Development of a Multiplex PCR Procedure for Rapid Discrimination between the Two Biotypes

Tararaj Dharakul,1 Boonratn Tassaneetrithep,1 Suwanna Trakulsomboon,2 and Sirirurg Songsivilai1,*

Laboratory of Cellular and Molecular Immunology, Department of Immunology,1 and Division of Infectious Disease, Department of Medicine,2 Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand

Received 9 October 1998/Returned for modification 8 December 1998/Accepted 17 March 1999

A Burkholderia pseudomallei-like organism has recently been identified among some soil isolates of B. pseudomallei in an area with endemic melioidosis. This organism is almost identical to B. pseudomallei in terms of morphological and biochemical profiles, except that it differs in ability to assimilate L-arabinose. These Ara+ isolates are also less virulent than the Ara- isolates in animal models. In addition, clinical isolates of B. pseudomallei available to date are almost exclusively Ara-. These features suggested that these two organisms may belong to distinctive species. In this study, the 16S rRNA-encoding genes from five clinical (four Ara- and one Ara+) and nine soil isolates (five Ara- and four Ara+) of B. pseudomallei were sequenced. The nucleotide sequences and phylogenetic analysis indicated that the 16S rRNA-encoding gene of the Ara+ biotype was similar to but distinctively different from that of the Ara- soil isolates, which were identical to the classical clinical isolates of B. pseudomallei. The nucleotide sequence differences in the 16S rRNA-encoding gene appeared to be specific for the Ara+ or Ara- biotypes. The differences were, however, not sufficient for classification into a new species within the genus Burkholderia. A simple and rapid multiplex PCR procedure was developed to discriminate between Ara- and Ara+ B. pseudomallei isolates. This new method could also be incorporated into our previously reported nested PCR system for detecting B. pseudomallei in clinical specimens.

* Corresponding author. Mailing address: Laboratory of Cellular and Molecular Immunology, Department of Immunology, Faculty of Medicine, Siriraj Hospital, Mahidol University, 2 Prannok Rd., Bangkok 10700, Thailand. Phone: 66-2-4197066. Fax: 66-2-4181636. E-mail: sissv{at}mahidol.ac.th.

Journal of Clinical Microbiology, June 1999, p. 1906-1912, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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