Coupled enzyme-based method for electronic monitoring of biological indicator
A sterilization indicator and microbial technology, applied in biochemical equipment and methods, microbial determination/inspection, biological testing, etc.
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Embodiment 1
[0082] For illustrative purposes, germinating spores of G. stearothermophilus at E5, E6 and E7 spore populations were monitored over several hours for each reaction containing the growth medium using a Biochemical Analyzer (YSI Instruments) using a glucose electrode. The growth medium consisted of glucose-free tryptone soy broth (TSB) at a concentration of 1 / 8 of the standard TSB concentration (in order to minimize interference from medium components) containing a concentration of 8.5 g / l or 10.5g / l complex sugar maltose. Glucose measurements were taken every 60 minutes. All samples were normalized to the initial glucose reading of the culture medium and observed over an extended time scale, measuring the overall kinetics of the response to the selected conditions. result in Figure 10 Shown in , which shows the effect of spore concentration and maltose concentration on the amount of glucose produced over time.
Embodiment 2
[0084] Evaluation of Geobacillus stearothermophilus spores under E5, E6, and E7 populations in various media formulations to determine how quickly glucose levels are produced when selected levels of maltose are broken down to glucose by viable germination organisms from the spores The presence of living organisms can be detected. The medium was prepared with different concentrations of glucose-free tryptone soy broth ranging from 1 / 8 of the standard glucose-free TSB (tryptone soy broth) concentration to the full standard concentration while keeping the maltose concentration constant at 8.5 g / l. Glucose-free TSB is a general nutrient source for bacteria and it contains casein tryptic digest, soybean papain digest, and salt and buffer; thus, by diluting it, we effectively reduce any interfering groups present in the culture amount of points. "Medium only" samples were monitored throughout the study in order to observe the effect of different concentrations of TSB on the backgr...
Embodiment 3
[0090] 12 containing 2x10 per SCBI 7 A self-contained biological indicator of Geobacillus stearothermophilus of the CPU population was placed in a simulated use medical device load and the full load was processed in a steam sterilizer in a failed (ie, incomplete) cycle. At the completion of the failed cycle, samples were removed and incubated at 56°C for 3 hours. Glucose levels in the medium were evaluated for each sample. Figure 12 is a graph showing the comparison between the signal measured when spores survived the sterilization treatment and the signal measured when no spores survived the sterilization treatment.
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