Conjugated enzyme-based method for electronic monitoring of biological indicators

一种灭菌指示器、微生物的技术，应用在生物化学设备和方法、微生物的测定/检验、生物测试等方向

Active Publication Date: 2017-04-05
AMERICAN STERILIZER CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Detection is limited by the need to generate signal (whether colorimetric or fluorescent) above basal background levels

Method used

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  • Conjugated enzyme-based method for electronic monitoring of biological indicators
  • Conjugated enzyme-based method for electronic monitoring of biological indicators
  • Conjugated enzyme-based method for electronic monitoring of biological indicators

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] For purposes of illustration, the Germination of G. stearothermophilus under E5, E6 and E7 spore populations was monitored over several hours for each reaction containing the growth medium using the Biochemical Analyzer (YSI Instruments) using a glucose electrode spores. 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

[0085] 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

[0091] 12 containing 2x 10 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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Abstract

A sterilization indicator system and method of determining the efficacy of a sterilization treatment using the system. The system comprises a bottle comprising a first compartment containing one or more microbial spores; a second compartment containing a growth medium comprising disaccharides, oligosaccharides or polysaccharides, wherein the bottle Contains no monosaccharides; enzymes capable of acting on said monosaccharides to produce reaction products and electron transfer, said enzymes being disposed on two or more electrodes adapted to carry the electrical signal of the electrode pair positioned to contact the contents of the merged first and second compartments during incubation; and a device connected or connectable to the electrodes and adapted to The electrical signal generated by electron transfer when the enzyme acts on monosaccharides is detected and measured.

Description

technical field [0001] The present invention relates to biological indicators for testing the efficacy of sterilization treatments, and more particularly to coupled enzyme-based methods for monitoring such biological indicators in which such monitoring can be performed electronically. Background technique [0002] Biological indicators (BI) are one of the most important types of indicators. Biological indicators provide the highest degree of assurance that the conditions for sterilization are met within the processor or by the load being processed itself. This type of biological indicator is intended to represent a worst-case scenario for a treatment system by providing within or on the indicator a very large number of organisms that are highly resistant to the particular treatment in question. Often bacterial spores are the organism of choice for monitoring sterilization systems. [0003] Biological indicators usually consist of microorganisms inoculated onto a carrier ma...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/22
CPCG01N33/52G01N2400/00Y10T436/25375Y10T436/11C12Q1/22C12Q1/006C12Q1/001G01N27/327
Inventor 菲利普·P·弗兰契什科维奇特里西娅·A·克雷格
Owner AMERICAN STERILIZER CO
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