| Budget Amount *help |
¥8,000,000 (Direct Cost: ¥8,000,000)
Fiscal Year 2000: ¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 1999: ¥3,700,000 (Direct Cost: ¥3,700,000)
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| Research Abstract |
(1) RAPID DETERMINATION OF ANTIMICROBIAL SUSCEPTIBILITY
The applicability of flow cytometry was studied to accelerate antimicrobial susceptibility testing. First, a new flow cytometer (MFCM) was developed for detection of as small particles as bacteria. MFCM measures the particle count, the pulse intensity and width of forward scattered light (Fsc and Fscw, respectively). Microsoft Excel software was used to analyze dual-parameter histogram (DPH) of Fsc vs. Fscw. Particle distributions on DPH were analyzed after incubation of bacterial solutions (12 species) with vigorous shaking at 37C in the presence and absence of serially diluted anatimicrobials (PIPC, LVFX, AMK and MINO). After incubation, PIPC-sensitive microbes shifted to the area with larger Fsc and Fscw, while LVFX-, AMK-and MINO-sensitive bacteria gathered to the area with smaller FSC. By comparing these consistent observations with the gold standard microdilution MICs, the quantitative criteria was developed for determining antimicrobial susceptibility within 2 hours, especially for gram-negative bacilli.
(2) RAPID DIAGNOSIS OF BACTERIURIA
In urinary tract infection, urine is often heavily contaminated with particles as small as bacteria. To differentiate bacteria with non-bacterial particles, urine samples were vigorously shaked for 1-2 hours at 37C and pre-incubation DPH was subtracted from post-incubation DPH. The net changes on subtracted DPH were indicative of bacterial growth. This method could contribute to the diagnosis of bacteriuria before microbes multiplied severalfold.
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