Rapid read-out biological indicator

• US 5,252,484 A
• Filed: 08/21/1991
• Issued: 10/12/1993
• Est. Priority Date: 11/29/1988
• Status: Expired due to Term

• Current Assignee: 3M Innovative Properties Company (3M Company)
• Original Assignee: Minnesota Mining and Manufacturing Co (3M Company)
• Inventors: Foltz, William E., Woodson, Lewis P., Matner, Richard R.
• Primary Examiner(s): KRATZ, PETER F
• Application Number:
  US07/748,327
• Time in Patent Office: 783 Days
• Field of Search: 435/31, 435/296, 435/288, 422/55, 422/58
• US Class Current: 435/287.4
• CPC Class Codes: C12Q 1/22 Testing for sterility conditions
• CPC Class Codes: Y10S 435/81 Packaged device or kit

Abstract

A rapid method of determining the efficacy of a sterilization cycle, and an indicator adapted to perform such method, comprising subjecting to the sterilization cycle a source of active enzyme having activity which correlates with the viability of a microorganism commonly used to monitor sterilization, and incubating the enzyme source, following the completion of the sterilization cycle, with an effective amount of a substrate system capable of reacting with any residual active enzyme to produce a detectable enzyme-modified product.

Claims

1. A sterility indicator comprising

a) an outer container having liquid impermeable and substantially gas non-absorptive walls, said container having at least one opening therein;

and

b) a gas-transmissive, bacterial impermeable means covering said opening; and

c) contained within said outer container, a detectable amount of active enzyme isolated from an appropriate microorganism, said enzyme having sufficient activity following a sterilization cycle which is sublethal to at least one test microorganism commonly used to monitor sterilization, to react with an effective amount of a substrate system for said enzyme to produce a detectable enzyme-modified product within less tan twenty-four hours, yet said enzyme having activity which is reduced to "background" following a sterilization cycle which is lethal to said test microorganisms.

2. The indicator of claim 1 further comprising in said outer container

d) an effective amount of an enzyme substrate capable of reacting in aqueous solution with active enzyme to produce a detectable enzyme-modified product.

3. The indicator of claim 2 wherein said enzyme substrate is in aqueous solution and is contained within a sealed, openable, gas and liquid impermeable inner container.

4. The indicator of claim 1 further comprising within the outer container a predetermined number of viable test microorganisms.

5. The indicator of claim 4 further comprising within the inner container an aqueous nutrient medium capable, with incubation, of promoting growth of said viable test microorganisms, and a detector material capable of undergoing a visible color change in response to growth of said test microorganism.

6. The sterility indicator of claim 1 wherein said enzyme is one which has sufficient activity following a sterilization cycle which is sublethal to at least one test microorganism commonly used to monitor sterilization, to react with said substrate system to produce a detectable enzyme-modified product within less than twelve hours.

7. The sterility indicator of claim 1 wherein said enzyme is one which has sufficient activity following a sterilization cycle which is sublethal to at least one test microorganism commonly used to monitor sterilization, to react with said substrate system to produce a detectable enzyme-modified product within less than eight hours.

8. The sterility indicator of claim 1 wherein said enzyme is one which has sufficient activity following a sterilization cycle which is sublethal to at least one test microorganism commonly used to monitor sterilization, to react with said substrate system to produce a detectable enzyme-modified product within less than three hours.

9. The indicator of claim 1 wherein said enzyme is one which

a) when subjected to a sterilization cycle which would be just sufficient to decrease the population of 1×

10⁶ test microorganisms to zero, as measured by lack of outgrowth of said microorganism, has activity equal to "background", as measured by reaction with an effective amount of any enzyme substrate system capable of reacting with said active enzyme to produce a detectable enzyme-modified product; and

b) when subjected to a sterilization cycle sufficient to decrease the population of 1×

10⁶ of said test microorganisms by at least about 1 log, but less than about 6 logs, has activity greater than said background, as measured by reaction with an effective amount of said enzyme substrate system.

10. The indicator of claim 1 wherein said enzyme is selected from the group consisting of beta-D-glucosidase, alpha-D-glucosidase, alkaline phosphatase, acid phosphatase, butyrate esterase, caprylate esterase lipase, myristate lipase, leucine aminopeptidase, valine aminopeptidase, chymotrypsin, phosphohydrolase, alpha-D-galactosidase, beta-D-galactosidase, alpha-L-arabinofuranosidase, beta-D-glucuronidase, N-acetyl-β

-glucosaminidase, beta-D-cellobiosidase, alanine aminopeptidase, proline aminopeptidase, tyrosine aminopeptidase, leucine aminopeptidase, phenylalanine aminopeptidase and a fatty acid esterase derived from spore-forming microorganisms.