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Gas chromatography–mass spectrometry method for rapid identification and differentiation of B urkholderia pseudomallei and B urkholderia mallei from each other, B urkholderia thailandensis and several members of the B urkholderia cepacia complex
Author(s) -
Li D.,
March J.K.,
Bills T.M.,
Holt B.C.,
Wilson C.E.,
Lowe W.,
Tolley H.D.,
Lee M.L.,
Robison R.A.
Publication year - 2013
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.12310
Subject(s) - burkholderia pseudomallei , burkholderia , microbiology and biotechnology , identification (biology) , mass spectrometry , biology , bacteria , chromatography , chemistry , genetics , botany
Abstract Aims To develop a simple gas chromatography–mass spectrometry ( GC ‐ MS ) method for the detection and differentiation of Burkholderia pseudomallei and Burkholderia mallei from each other, Burkholderia thailandensis and several members of the Burkholderia cepacia complex. Methods and Results Biomarkers were generated by one‐step thermochemolysis ( TCM ) and analysed using a GC ‐ MS system. Fragments of poly‐3‐hydroxybutyrate‐ co ‐hydroxyvalerate [poly(3 HBA ‐ co ‐3 HVA )] produced by TCM were useful biomarkers. Several cellular fatty acid methyl esters were important in differentiating the various Burkholderia species. A statistical discrimination algorithm was constructed using a combination of biomarkers. The identities of four B. pseudomallei strains, four B. mallei strains and one strain of each near neighbour were confirmed in a statistically designed test using the algorithm. The detection limit for this method was found to be approximately 4000 cells. Conclusions The method is fast, accurate and easy to use. The algorithm is robust against different growth conditions (medium and temperature). Significance and Impact of the Study This assay may prove beneficial in a clinical diagnostic setting, where the rapid identification of B. pseudomallei is essential to effective treatment. This method could also be easily employed after a biological attack to confirm the presence of either B. pseudomallei or B. mallei .