J Bacteriol Virol.  2011 Mar;41(1):19-25. 10.4167/jbv.2011.41.1.19.

The Laboratory Diagnosis of Melioidosis in a Korean Patient

• Shin YW
• Cho MH
• Chun JH
• Kim C
• Oh HB
• Rhie GE
• Yoo CK

• KMID: 1449859
• DOI: http://doi.org/10.4167/jbv.2011.41.1.19

Abstract

Burkholderia pseudomallei is a gram-negative opportunistic intracellular pathogen that causes an acute and fatal septicemic melioidosis in humans. The organism is mainly found in Southeastern Asia and Northern Australia. Recently, we encountered a case of melioidosis in a Korean patient and performed the laboratory diagnosis of melioidosis. As a result, a gram negative bacterium was isolated from a melioidosis patient, and it was identified as B. pseudomallei on DNA sequencing of 16S ribosomal RNA with 99.9% homology and biochemical examination of VITEK gram-negative identification card. Also, DNA from cultured bacteria was tested in multiplex PCR, a 245 bp fragment amplified from the metalloprotease gene and a fragment of variable size ranging from 400~700 bp resulting from amplification of the 10 bp repetitive element for B. pseudomallei were confirmed after electrophoresis. The bacterium was sensitive to ceftazidime, imipenem and meropenem but resistant to ticarcillin. So far, there are no domestic cases of melioidosis in Korea, however, due to the increase in international travelers, the incidence of melioidosis is likely to increase. We report a recent case of melioidosis in a Korean patient.

Keyword

Burkholderia pseudomallei; Melioidosis; Diagnosis; VITEK; Multiplex PCR

MeSH Terms

Asia, Southeastern
Australia
Bacteria
Burkholderia pseudomallei
Ceftazidime
Clinical Laboratory Techniques
DNA
Electrophoresis
Humans
Imipenem
Incidence
Korea
Melioidosis
Multiplex Polymerase Chain Reaction
RNA, Ribosomal, 16S
Sequence Analysis, DNA
Thienamycins
Ticarcillin
Ceftazidime
DNA
Imipenem
RNA, Ribosomal, 16S
Thienamycins
Ticarcillin

Figure

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  Figure 1. Colonial growth morphology of Burkholderia pseudomallei. (A) Colonies on BHI agar after 48 h culture; (B) Colonies on MacConkey agar after 48 h culture; (C) Colonies on Ashdown's medium after 48 h culture.

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  Figure 2. Comparison of the 16S rRNA sequences from B. pseudomallei (GenBank accession no. CP000572) and B. pseudomallei isolated from a melioidosis patient. Bold letters show the positions of the 3 nucleotide dissimilarities between B. pseudomallei and B. pseudomallei isolated from a melioidosis patient.

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  Figure 3. The phylogenetic tree based on the 16S rRNA gene sequences for B. pseudomallei isolate and other Burkholderia. The phylogenetic tree was generated using Neighbor-joining method based on DNA sequence of partial 16S rDNA gene. Bootstrap values of above 50% from a sample of 1,000 replicate are shown on each branch.

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  Figure 4. Multiplex PCR analysis of B. mallei, B. thailandensis and B. pseudomallei isolates. Lane 1. B. mallei; lane 2. B. thailandensis; lane 3. B. pseudomallei; lane 4. Genomic DNA isolated directly from blood; lane 5. B. pseudomallei cultured from blood; lane 6. B. pseudomallei cultured from sputum; lane 7. B. cepacia; lane 8. P. aeruginosa; lane 9. E.coli.

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