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Development of a novel method for detection of Clostridium difficile using HS-SPME-GC-MS

Lookup NU author(s): Dr John Perry



AimsA novel method has been developed that allows successful differentiation between Clostridium difficile culture-positive and culture-negative stool samples based on volatile organic compound (VOC) evolution and detection by headspace solid-phase microextraction coupled with gas chromatography mass spectrometry (HS-SPME-GC-MS).Methods and ResultsThe method is based on the activation of p-hydroxyphenylacetate decarboxylase produced by Cl.difficile and the detection of a specific VOC, that is 2-fluoro-4-methylphenol from an enzyme substrate. In addition, other VOCs were good indicators for Cl.difficile, that is isocaproic acid and p-cresol, although they could not be used alone for identification purposes. One hundred stool samples were tested, of which 77 were positive by culture. Detection using HS-SPME-GC-MS allowed confirmation of the presence of Cl.difficile within 18h with a sensitivity and specificity of 83 center dot 1 and 100%, respectively.ConclusionsIt is recommended that this new approach could be used alongside conventional methods for Cl.difficile detection, including toxin detection methods, which would allow any false-negative results to be eliminated.Significance and Impact of the StudyThe ability to identify Cl.difficile-positive stool samples by the analysis of VOCs could allow the development of a VOC detection device which could allow rapid diagnosis of disease and hence prompt treatment with appropriate antibiotics.

Publication metadata

Author(s): Tait E, Hill KA, Perry JD, Stanforth SP, Dean JR

Publication type: Article

Publication status: Published

Journal: Journal of Applied Microbiology

Year: 2014

Volume: 116

Issue: 4

Pages: 1010-1019

Print publication date: 06/01/2014

Date deposited: 08/08/2014

ISSN (print): 1364-5072

ISSN (electronic): 1365-2672

Publisher: Wiley-Blackwell


DOI: 10.1111/jam.12418


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