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Journal of Clinical Microbiology, May 2005, p. 2201-2206, Vol. 43, No. 5
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.5.2201-2206.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Comparison of Diagnostic Laboratory Methods for Identification of Burkholderia pseudomallei

Timothy J. J. Inglis,* Adam Merritt, Glenys Chidlow, Max Aravena-Roman, and Gerry Harnett

Division of Microbiology & Infectious Diseases, Western Australian Centre for Pathology and Medical Research (PathCentre), Nedlands, Western Australia 6009, Australia

Received 24 November 2004/ Returned for modification 30 December 2004/ Accepted 4 January 2005

Limited experience and a lack of validated diagnostic reagents make Burkholderia pseudomallei, the cause of melioidosis, difficult to recognize in the diagnostic microbiology laboratory. We compared three methods of confirming the identity of presumptive B. pseudomallei strains using a collection of Burkholderia species drawn from diverse geographic, clinical, and environmental sources. The 95 isolates studied included 71 B. pseudomallei and 3 B. thailandensis isolates. The API 20NE method identified only 37% of the B. pseudomallei isolates. The agglutinating antibody test identified 82% at first the attempt and 90% including results of a repeat test with previously negative isolates. Gas-liquid chromatography analysis of bacterial fatty acid methyl esters (GLC-FAME) identified 98% of the B. pseudomallei isolates. The agglutination test produced four false positive results, one B. cepacia, one B. multivorans, and two B. thailandensis. API produced three false positive results, one positive B. cepacia and two positive B. thailandensis. GLC-FAME analysis was positive for one B. cepacia isolate. On the basis of these results, the most robust B. pseudomallei discovery pathway combines the previously recommended isolate screening tests (Gram stain, oxidase test, gentamicin and polymyxin susceptibility) with monoclonal antibody agglutination on primary culture, followed by a repeat after 24 h incubation on agglutination-negative isolates and GLC-FAME analysis. Incorporation of PCR-based identification within this schema may improve percentages of recognition further but requires more detailed evaluation.


* Corresponding author. Mailing address: Division of Microbiology & Infectious Diseases, PathCentre, Locked Bag 2009, Nedlands, WA 6009, Australia. Phone: 618 9346 3461. Fax: 618 9381 7139. E-mail: tim.inglis{at}health.wa.gov.au.


Journal of Clinical Microbiology, May 2005, p. 2201-2206, Vol. 43, No. 5
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.5.2201-2206.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.




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