Biologically relevant methods for the rapid determination of sterility

A microorganism-free process for determining the microorganism survivability of a sterilizing environment comprising the steps of:

• a) exposing one or more components, but not all of the components of a plurality of interactive enzyme systems to a sterilizing environment;
• b) adding the remaining components of the plurality of interactive enzyme systems to the one or more components which were subjected to the sterilizing environment;
• and
• c) detecting activity of the plurality of interactive enzyme systems to determine the microorganism survivability of the sterilizing environment.

A microorganism-free process for determining the microorganism survivability of a sterilizing environment comprising the steps of:

• a) exposing an inhibitor of a plurality of interactive enzyme systems to a sterilizing environment;
• b) adding the remaining components of the plurality of interactive enzyme systems to the inhibitor which was subjected to the sterilizing environment;
• and
• c) detecting the activity of the plurality of interactive enzyme systems to determine the microorganism survivability of the sterilizing environment.

A two-step biological process for determining the effectiveness of a sterilization procedure comprising the steps of:

• a) placing a test indicator containing one or more components, but not all of the components of a plurality of interactive enzyme systems and a sample of microorganisms into a sterilization chamber;
• b) performing the sterilization procedure within the chamber;
• c) removing the test indicator from the chamber;
• d) adding the remaining components of the plurality of interactive enzyme systems to the one or more components in the test indicator to form a mixture and adding a fluid media to the sample of microorganisms to form a culture;
• e) incubating the mixture for a first period of time sufficient for detecting the presence or absence of a detectable product to make a first-step determination of the effectiveness of the sterilization procedure;
• and
• f) incubating the culture for a second period of time sufficient for detecting the presence or absence of a metabolic product of growth to make a second-step determination of the effectiveness of the sterilization procedure.

A self-contained test indicator for determining the effectiveness of a sterilization procedure comprising:

• a) an outer container having liquid impermeable and substantially gas non-absorptive walls and at least one opening covered with a gas-transmissive cover, said at least one opening leading into a chamber that contains a first and a second vial wherein;
• b) the first vial has at least one opening which is covered with a gas-transmissive cover and contains at least one component but not all of the component of a plurality of interactive enzyme systems;
• and
• c) the second vial contains the remaining components of the plurality of interactive enzyme systems which when mixed with the at least one component of the first vial produces a detectable product wherein;
• d) said vials are constructed of a material which can be opened and their contents mixed without being removed from the test indicator.

A biological process for determining the effectiveness of a sterilization procedure comprising the steps of:

• a) placing a test indicator containing one or more component, but not all of the components of a plurality of interactive enzyme systems into a sterilization chamber;
• b) performing the sterilization procedure within the chamber;
• c) adding the remaining components of the plurality of interactive enzyme systems to the one or more components in the test indicator to form a mixture;
• d) incubating the mixture for less than one minute;
• and
• e) detecting the presence or absence of a product.

A biological process for determining the effectiveness of a sterilization procedure comprising the steps of:

• a) placing a test indicator containing one or more components but not all of the components of a plurality of interactive enzyme systems into a sterilization chamber;
• b) performing the sterilization procedure within the chamber;
• c) adding the remaining components of the plurality of interactive enzyme systems to the one or more components in the test indicator;
• and
• d) detecting the presence or absence of a product.

A test kit for determining the effectiveness of a sterilization procedure comprising:

• a) a first container having liquid impermeable and substantially gas non-absorptive walls that is to be subjected to a sterilizing environment;
• b) at least one opening in the container leading into a chamber which contains at least two components, but not all the components, of a plurality of interactive enzyme systems, with a gas-transmissive barrier between the at least two components and the at least one opening;
• and
• c) a second container containing the remaining components of the plurality of interactive enzyme systems which are to be added to the at least two or more components that are subjected to the sterilizing environment.