The use of high-resolution 1H nuclear magnetic resonance (NMR) spectroscopy in the clinical diagnosis of Acanthamoeba

Acanthamoeba are opportunistic protozoan pathogens that can produce sight-threatening keratitis and fatal granulomatous encephalitis. The successful prognosis requires early diagnosis and differentiation of pathogenic Acanthamoeba spp. followed by aggressive treatment regimen. In this study, we tested the use of high-resolution H-1 NMR spectroscopy in the clinical diagnosis of Acanthamoeba. Using NMR spectroscopy combined with Pattern Recognition Analysis (PRA), we analysed variations in the biochemical 'fingerprint' of invasive and non-invasive Acanthamoeba, its closely related genus, Balamuthia mandrillaris, neuropathogenic Escherichia coli K1 strain E44, a laboratory strain of E. coli K-12, HB101, mammalian cells including human brain microvascular endothelial cells and monkey kidney cells. The findings revealed significant variations in the metabolites of amoebae, mammalian cells and bacteria. Interestingly, H-1 NMR spectra provided distinct biochemical profiles of clinical and non-clinical Acanthamoeba isolates highlighting the potential of H-1 NMR spectroscopy combined with PRA for the development of a novel diagnostic test that could rapidly identify pathogenic Acanthamoeba isolates with high sensitivity and specificity.