Detecting bacterial lung infections: in vivo evaluation of in vitro volatile fingerprints

Abstract

The identification of bacteria by their volatilomes is of interest to many scientists and clinicians as it holds the promise of diagnosing infections in situ, particularly lung infections via breath analysis. While there are many studies reporting various bacterial volatile biomarkers or fingerprints using in vitro experiments, it has proven difficult to translate these data to in vivo breath analyses. Therefore, we aimed to create secondary electrospray ionization-mass spectrometry (SESI-MS) pathogen fingerprints directly from the breath of mice with lung infections. In this study we demonstrated that SESI-MS is capable of differentiating infected versus uninfected mice, P. aeruginosa-infected versus S. aureus-infected mice, as well as distinguish between infections caused by P. aeruginosa strains PAO1 versus FRD1, with statistical significance (p < 0.05). In addition, we compared in vitro and in vivo volatiles and observed that only 25-34% of peaks are shared between the in vitro and in vivo SESI-MS fingerprints. To the best of our knowledge, these are the first breath volatiles measured for P. aeruginosa PAO1, FRD1, and S. aureus RN450, and the first comparison of in vivo and in vitro volatile profiles from the same strains using the murine infection model.

Fig. 1

SESI-MS breathprints of mice with P. aeruginosa PAO1, P. aeruginosa FRD1, or S. aureus RN450 lung infections, or uninfected lungs. Each spectrum is the average of breath from five mice after a 24 h lung infection.

Figure 2

SESI-MS spectra of P. aeruginosa PAO1, FRD1, and S. aureus RN450, grown in vitro in TSB (24 h, 37°C)