Analysis of methicillin-resistant Staphylococcus aureus isolates by proton magnetic resonance spectroscopy.
OBJECTIVE
To evaluate the stability and(1)H nuclear magnetic resonance ((1)H NMR) differences among bacterial strains, we analysed(1)H NMR spectra for 50 methicillin-resistant Staphylococcus aureus (MRSA) isolates from 42 patients.


METHODS
(1)H NMR spectra for 50 MRSA isolates were obtained at 30 degrees C using a JNM-GX 270 NMR spectrometer at a field strength of 6.34 Tesla (270 MHz for(1)H). DNA fingerprints were obtained by pulsed-field gel electrophoresis (PFGE).


RESULTS
Each strain had eight to nine specific resonance features in the 0.5-4.5 ppm spectral region. These features were found in all strains, but the intensity of each feature varied between strains. Six resonance features (A-F) were selected for investigation. The relative integrated intensity (mean+/-SD, normalized to feature C = 1) of each feature was: feature A, 0.49+/-0.16; feature B, 3.93+/-0.81; feature D, 0.38+/-0.22; feature E, 1.51+/-0.96; feature F, 2.18+/-0.47. Within-strain reproducibility of feature intensities for A, B, D and F was good (coefficient of variation < 10%) for replicate cultures, analyzed on separate days. Feature E showed poor within-strain reproducibility. Storage at 4 degrees C for 4 months or disintegration of the micro-organisms by ultrasound did not alter(1)H NMR spectra. Two isolates from different patients, but the same hospital, showed indistinguishable NMR spectra, and were also indistinguishable in PFGE. Five strains with distinct PFGE patterns showed differences in NMR spectra outside the range of within-strain variation.


CONCLUSIONS
It is possible to analyse the whole molecular structure of MRSA by(1)H NMR. With this technique, we established that there are detectable, reproducible differences in quantitative cell composition between MRSA strains.(1)H NMR spectroscopy appears to have potential as a useful tool for epidemiological typing of bacteria.
Ohara
T
T
Department of Clinical Pathology, Jichi Medical School, Tochigi Prefecture 329-0498, Japan.
Itoh
Y
Y
Itoh
K
K
Tetsuka
T
T
D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D016521:Electrophoresis, Gel, Pulsed-Field; D006801:Humans; D016106:Methicillin Resistance; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011522:Protons; D015203:Reproducibility of Results; D013211:Staphylococcus aureus / Q000737:chemistry* / Q000145:classification / Q000302:isolation & purification
D004269:DNA, Bacterial; D011522:Protons