| Budget Amount *help |
¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1999: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1998: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1997: ¥500,000 (Direct Cost: ¥500,000)
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| Research Abstract |
First we applied proton nuclear magnetic resonance (ィイD11ィエD1H NMR) spectroscopy to distinguish between four bacterial species : Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosae and Kiebsiella pneumoniae. Because of differences in the composition of their cell wall and cytoplasmic constituents, individual bacterial species exhibited characteristic spectra except for K pneumoniae. Reproducibility was satisfactory in the quantitative studies for chemical shift, intensity of signal and integrated intensity. The turnaround time of the test was only about 40 minutes. ィイD11ィエD1H NMR could be a new powerful tool for bacterial identification.
Moreover, we analyzed ィイD11ィエD1H NMR spectra for 50 methicillin-resistant Staphylococcus aureus (MRSA) isolates from 42 patients to evaluate the stability and the ィイD11ィエD1H NMR differences among bacterial strains. Each strain had a reproducible, characteristic NMR spectral pattern containing eight to nine specific resonance or groups of re
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sonances in the 0.5 to 4.5 ppm spectral region. Six resonance groups were identified according to the resonance of the highest peak (group A - F). The relative integrated intensity (mean ± SD, normalized to group C=1) of each group was 0.49 ± 0.16 (group A), 3.93 ± 0.81 (group B), 0.38 ± 0.22 (group D), 1.51 ± 0.96 (group E) and 2.18 ± 0.47 (group F). Regarding to reproducibility of the NMR spectra, the C.V. was less than 10% in all groups except group E. The configuration of each NMR spectral group among replicates of the four strains used in the reproducibility was almost identical with only minor variations in the demonstrated intensity of resonances. Storage at 4°C for 4 months or destruction of the microorganisms by ultrasound did not alter ィイD11ィエD1H NMR spectra. NMR spectra of two MRSA strains that had identical DNA patterns by the restriction enzyme were similar. The correlation coefficient between the two NMR spectra was 0.998. Thus, ィイD11ィエD1H NMR spectroscopy appears to be a useful tool for strain identification. Less
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