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. 2010 Nov 7:10:56.
doi: 10.1186/1471-2466-10-56.

2-Aminoacetophenone as a potential breath biomarker for Pseudomonas aeruginosa in the cystic fibrosis lung

Amy J Scott-Thomas  1 , Mona Syhre, Philip K Pattemore, Michael Epton, Richard Laing, John Pearson, Stephen T Chambers
Affiliations

2-Aminoacetophenone as a potential breath biomarker for Pseudomonas aeruginosa in the cystic fibrosis lung

Amy J Scott-Thomas et al. BMC Pulm Med. .

Abstract

Background: Pseudomonas aeruginosa infections are associated with progressive life threatening decline of lung function in cystic fibrosis sufferers. Growth of Ps. aeruginosa releases a "grape-like" odour that has been identified as the microbial volatile organic compound 2-aminoacetophenone (2-AA).

Methods: We investigated 2-AA for its specificity to Ps. aeruginosa and its suitability as a potential breath biomarker of colonisation or infection by Solid Phase Micro Extraction and Gas Chromatography-Mass Spectrometry (GC/MS).

Results: Cultures of 20 clinical strains of Ps. aeruginosa but not other respiratory pathogens had high concentrations of 2-AA in the head space of in vitro cultures when analysed by GC/MS. 2-AA was stable for 6 hours in deactivated glass sampling bulbs but was not stable in Tedlar® bags. Optimisation of GC/MS allowed detection levels of 2-AA to low pico mol/mol range in breath. The 2-AA was detected in a significantly higher proportion of subjects colonised with Ps. aeruginosa 15/16 (93.7%) than both the healthy controls 5/17 (29%) (p < 0.0002) and CF patients not colonised with Ps. aeruginosa 4/13(30.7%) (p < 0.001). The sensitivity and specificity of the 2-AA breath test compared to isolation of Ps. aeruginosa in sputum and/or BALF was 93.8% (95% CI, 67-99) and 69.2% (95% CI, 38-89) respectively. The peak integration values for 2-AA analysis in the breath samples were significantly higher in Ps. aeruginosa colonised subjects (median 242, range 0-1243) than the healthy controls (median 0, range 0-161; p < 0.001) and CF subjects not colonised with Ps. aeruginosa (median 0, range 0-287; p < 0.003).

Conclusions: Our results report 2-AA as a promising breath biomarker for the detection of Ps. aeruginosa infections in the cystic fibrosis lung.

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Figures

Figure 1
Figure 1
NIST library formula and mass spectra of 2-AA under EI conditions.
Figure 2
Figure 2
Experimental spectra for 2-AA, obtained under GC/MS EI conditions, from in vitro Ps. aeruginosa cultures.
Figure 3
Figure 3
Experimental spectra for 2-AA, obtained under GC/MS-MS CI conditions, from in vitro Ps. aeruginosa cultures.
Figure 4
Figure 4
Experimental spectra for 2-AA, obtained under GC/MS3 CI conditions, from a breath samples positive for 2-AA.
Figure 5
Figure 5
A time degradation graph of 2-AA in glass sampling bulbs. The 2-AA levels (mean and SD) from three replicate experiments are shown and a five parameter logistic function fitted to the degradation curve (11.73 hours (95% CI 10.94, 12.52). 2-AA levels at 8 and 10 hours were significantly less (P < 0.001) than four and six hours.
Figure 6
Figure 6
2-AA detection in breath. The peak integration values for 2-AA were significantly higher in CF patients colonised with Ps. aeruginosa than in CF patients not colonised (p < 0.001) with Ps. aeruginosa and healthy subjects (p < 0.0002). The numbers of subjects with detectable 2-AA in each group are shown. The lower limit of detection (peak integration of 88) is shown as a dotted line.
See this image and copyright information in PMC

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