title: Comparison of diagnostic laboratory methods for identification of Burkholderia pseudomallei.
abstract: 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.
authors.affiliation: Division of Microbiology & Infectious Diseases, Western Australian Centre for Pathology and Medical Research (PathCentre), Locked Bag 2009, Nedlands, WA 6009, Australia. tim.inglis@health.wa.gov.au
authors.forename: Timothy J J
authors.initials: TJ
authors.lastname: Inglis
authors.forename: Adam
authors.initials: A
authors.lastname: Merritt
authors.forename: Glenys
authors.initials: G
authors.lastname: Chidlow
authors.forename: Max
authors.initials: M
authors.lastname: Aravena-Roman
authors.forename: Gerry
authors.initials: G
authors.lastname: Harnett
mesh_terms: D000818:Animals; D001483:Base Sequence; D016957:Burkholderia pseudomallei / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D002849:Chromatography, Gas / Q000379:methods; D017931:DNA Primers; D005227:Fatty Acids / Q000032:analysis; D006801:Humans; D008554:Melioidosis / Q000175:diagnosis*; D016133:Polymerase Chain Reaction; D014904:Western Australia
chemical_list: D017931:DNA Primers; D005227:Fatty Acids