Sterility indicating biological compositions, articles and methods

A technology of sterilization indication and composition, which is applied in the fields of biochemical equipment and methods, enzyme production/bioreactor, and microorganism determination/inspection, etc., and can solve problems such as excessive time

Inactive Publication Date: 2011-11-16
3M INNOVATIVE PROPERTIES CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although progress has been made, the time required for definitive measurements may be too long

Method used

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  • Sterility indicating biological compositions, articles and methods
  • Sterility indicating biological compositions, articles and methods
  • Sterility indicating biological compositions, articles and methods

Examples

Experimental program
Comparison scheme
Effect test

example

[0171] Spore suspension

example 1

[0174] Protease detection

[0175] Dilutions of the spore suspension of G. stearothermophilus were made in sterile rinse water (Baxter, Deerfield, IL). The resulting diluted spore preparation (1 μL) was diluted with 1×10 6 spores, 1×10 5 spores, 1×10 4 spores, 1×10 3 The final population of 0 and 0 spores was assayed in a plate reader as described below. The spores were dried for 20 minutes in a 37°C incubator. The spores were then rehydrated in 100 uL of medium consisting of: 50 uL of GFK (1 mg / mL glucose, 1 mg / mL fructose, 3.3 mg / mL potassium chloride), 25 uL of Tris buffer (containing 0.2 mM CaCl) and 25 uL of 4 mg / mL labeled protease substrate (Calbiochem, casein labeled with N-(resorufin-4-carbonyl)piperidine-4-carbonic acid, protease substrate (EMD Chemicals, Gibbstown, NJ) ). Each resulting mixture was added to the wells of the plate. Plates were then placed in a preheated Synergy 4 plate reader (BioTech, Winooski, VT) at 50°C and incubated for 480 minutes. D...

example 2

[0180] Stability of Labeled Protease Substrates to Exposure to Sterilization Temperatures

[0181] Dilutions of the spore suspension of G. stearothermophilus were made in sterile rinse water (Baxter, Deerfield, IL). The resulting diluted spore preparation (1 μL) was diluted with 1×10 6 The final populations of spores and 0 spores were tested. The spores were dried for 20 minutes in a 37°C incubator. The spores were then rehydrated in 100 uL of medium consisting of: 50 uL of GFK (1 mg / mL glucose, 1 mg / mL fructose, 3.3 mg / mL potassium chloride), 25 uL of Tris buffer (containing 0.2 mM CaCl) and 25uL of the 4mg / mL labeled protease substrate as in Example 1, the culture medium was previously carried out at 121° C. for 15 minutes in an AMSCO Scientific SG-120 Eagle / Century Series steam sterilizer (Steris, Mentor, OH). (250°F) vacuum assisted circulation. At the same time, a culture medium not subjected to sterilization conditions (non-autoclaved protease medium) was tested as...

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Abstract

The present invention discloses a sterility indicating composition comprising a plurality of sterilization process resistant spores which contain an active protease during germination and initial outgrowth of the spores; and a germination medium comprising at least one labeled protease substrate and at least one nutrient for germination of the spores; wherein the medium is essentially free of a) any active protease other than the active protease contained by the plurality of spores and b) any protease substrate other than the at least one labeled protease substrate, other than any protease substrate originating from the plurality of spores, and other than any protease substrate which does not compete with the labeled protease substrate for the active protease; and wherein the at least one labeled protease substrate comprises a peptide which can be cleaved by the active protease and which is labeled with one or more dye groups, at least one of which undergoes a detectable change when the peptide is cleaved by the active protease, and wherein the labeled protease substrate is stable at least at a temperature for incubating the spores, a sterilization process indicator comprising the composition, and a method of determining the effectiveness of a sterilization process using the composition and indicator are disclosed.

Description

[0001] Cross references to related patent applications [0002] This application claims the benefit of US Provisional Application No. 61 / 196414, filed October 17, 2008, which is hereby incorporated by reference in its entirety. Background technique [0003] Primarily in the healthcare industry, but also in many other industrial applications, it is necessary to monitor the effectiveness of processes used to sterilize equipment such as medical devices, instruments and other non-disposable items. In these contexts, sterilization is generally defined as the process of complete destruction of all living microorganisms, including structures such as viruses and spores. As standard practice, hospitals place a sterilization status indicator with a batch of items to test the lethality of the sterilization process. Both biological and chemical sterilization indicators have been utilized. [0004] Standard types of biological sterilization indicators include known quantities of test mi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L2/28C12Q1/18C12M1/40
CPCA61L2/28C12Q1/22C12Q1/37
Inventor 塞拉嘉·常德拉帕蒂希瑟·M·韦布魏爱平
Owner 3M INNOVATIVE PROPERTIES CO
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