3M Infection Prevention Solutions

3M Sterile U Web Meeting July 17, 2014

The Science of Speed The Evolution of Biological Indicators

Welcome!
Topic: The Science of Speed The Evolution of Biological Indicators
Facilitators:

Christophe de Campeau, 3M

Sandra Velte, 3M

Speakers: Craig Wallace Senior Technical Specialist 3M

Housekeeping

Questions

Todays meeting times are

9:00 a.m., 11:00 a.m. and 1:00 p.m. CST

How do I get a CE Certificate?

difficulties
Craig Wallace 3M

follow-up

For22 more information: www.3M.com/3MSterileU

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Learning Objectives

Next week, all of todays meeting participants will be

sent an email containing instructions for obtaining a
CE Certificate for todays meeting.

1. Describe the design and function of biological indicators

2. Discuss how biological indicator incubation time is
determined

The email will be sent to the email address you

provided when you logged-in to todays meeting. If
there are others listening with you today who did not
log-on, you may forward the CE certificate email to
them.

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Chat feature

Post session
2

Mute feature (*7 = unmute; *6 = mute)

Technical

To hear audio, call 800-937-0042 and enter access code 7333633

Phone lines are muted. Audio will commence when the webinar begins.

Sterilization Monitoring - Quality Control

3. Understand how rapid readout biological indicator systems

work

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Sterilization Process Monitoring Tools

Load
Control

Biological
Indicators

Pack
Control
Equipment
Control

Chemical
Indicators

Physical
Monitors

Exposure
Control

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Late 1800s . . .

The history of biological indicators

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First biological indicators . . .

National Institutes of Health (NIH) 1955

Recommends Bacillus stearothermophilus* as the test organism for
steam sterilization

Robert Koch 1881

Forms spores

No threat

- challenges process

of pathogenicity, pyrogenicity, or toxicity

Inoculated carriers
* Now known as
Geobacillus
stearothermophilus

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The basic principle of biological indicators

Direct measurement

Large

BI Basics . . . .

of lethality

number of test organisms

Test organisms

that are very resistant to the

process (hard to kill)
If the process can kill the BI, it will kill the lower
numbers of less resistant organisms on the
medical devices

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Bacterial Spores

Bacterial Spores
Some types of bacteria can form spores to
protect themselves from a hostile environment,
such as lack of nutrients

e.g. some

Bacillus sp., Clostridium sp.

Protective

shell around DNA, almost no water

No biological

activity - dormant

Geobacillus
stearothermophilus

Google Images

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Bacterial Spores - Formation

Bacillus
atropheus

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Bacterial Spores Activation and germination

Many complex steps driven by enzymes

Google Images
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Bacterial Growth
Bacterial Growth

If we had a starting count of 10 contaminating

bacteria that can grow and double every
20 minutes, you will have:
Elapsed Time
0:00
0:20
0:40
1:00
1:20
1:40
2:00
2:20
2:40
3:00
3:20
3:40
4:00
4:20
4:40
5:00
5:20
5:40
6:00

Google Images

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Types of BIs Inoculated devices

# of Bacteria
10
20
40
80
180
360
720
1,440
2,880
5,760
11,520
23,040
46,080
92,160
184,320
368,640
737,280
1,474,560
2,949,120

Cycle time:
20 minutes
- 3 hours
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Types of BIs Spore Strips

Types of BIs Spore Ampoules

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Types of BIs Self-contained (SCBI)

Self-contained BIs color change

Growth of the test organisms produces acid

pH of the growth media drops

Filter

Cap

Growth
Media
with pH
Indicator

Spore
Strip

pH indicator changes color positive BI

Sleeve
Negative

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Positive

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Biological indicator testing - resistometer

Biological
indicator
performance
requirements
ANSI/AAMI/ISO
11138 series

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Key performance parameters

Biological indicators key specifications

Population (number of test organisms)

Number

Moist heat

of organisms that must be killed to get a negative BI

The more

organisms, the longer it takes to inactive the BI

Measured

by distributing the spores on plates and counting

Expressed

in exponents (e.g. 106 = 1,000,000)

D-value:

D-value (resistance)

A measure

1.5 minute minimum (121C)

Population:

1 x 105 CFU (minimum)

Ethylene oxide

of how resistant the organisms are to the process (how hard to kill)

Measured

as the time to reduce the population by 90%

D-value:

Expressed

in time units (usually minutes)

Population:

2.5 minutes minimum

1 x 106 CFU minimum

ISO 11138:2006
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The evolution of BI technology

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Biological indicator readout time

BI evolution
Consistency

Purified cultures

Readout

(or incubation) time: Time required to

decide that a biological indicator is NEGATIVE

of carefully selected test organisms

7 days is the accepted

readout time for BIs for

established sterilization processes, with known
test organisms

Convenience

Self-contained

designs eliminate transfers to growth

media

Readout time

7 days

reduced to minutes
ISO/ANSI/AAMI 11138-1: 2006

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FDA Guidance
Reduced
Incubation Time

Reduced incubation time (RIT)

A BI readout time of less than 7 days, which has been

validated to an appropriate level of accuracy as

compared to the 7 day result.

standard seven or
more days

How is that done?

more than 97% . . .

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Evolution of readout times

Rapid biological indicator technologies*
?????

7 days

24 - 48 hours

1-4 hours

30 - 60 minutes
*Less than 24 hours
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Bacterial Growth
Bacterial Growth

If we had a starting count of 10 contaminating

bacteria that can grow and double every
20 minutes, you will have:
Elapsed Time
0:00
0:20
0:40
1:00
1:20
1:40
2:00
2:20
2:40
3:00
3:20
3:40
4:00
4:20
4:40
5:00
5:20
5:40
6:00

Mesa Labs Smart-Read Technology

# of Bacteria
10
20
40
80
180
360
720
1,440
2,880
5,760
11,520
23,040
46,080
92,160
184,320
368,640
737,280
1,474,560
2,949,120

Standard SCBI growth

media color change

Colorimeter reads color

shift and declares a positive when certain

color change criteria are met

Readout time

Cycle time:
20 minutes
- 3 hours
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is 10 hours

Smart-Read is a registered trademark of Mesa

Labs, Inc.
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www.mesalabs.com

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Bacterial spores activation and

germination

Spore start up . . . Computer start up

Many complex steps driven by enzymes

Google Images
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3M Rapid Attest and Super Rapid Attest SCBIs

Technology: 3M Rapid Attest and

Super Rapid Attest Technology
Detects biological activity very early in the
spores activation and germination process

Takes advantage of the activity of enzymes used by the cell

Super Rapid Attest

Cap

to germinate and grow

Detects the end product of the enzyme reactions

Growth
Media with
pH Indicator
and
fluorescent
indicator

Spore
carrier

Sleeve

Glucosidase enzymes - carbohydrate metabolism

Geobacillus stearothermophilus: alpha-glucosidase
Bacillus atrophaeus: beta-glucosidase

Rapid Attest

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3M Rapid Readout Technology

3M Rapid Readout Technology

Non-fluorescent substrate
in growth media

Non-fluorescent substrate
in growth media

Fluorescent by-product (4-MU)

detected in Auto-reader

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Fluorescent by-product (4-MU)

detected in Auto-reader

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Readout Times

Rapid Attest and Super Rapid Attest also

can provide a conventional outgrowthbased pH color change response

Rapid

Attest 1292

3 hours

Super

Rapid Attest 1492V

1 hour

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Summary
Biological indicators provide a direct microbial
challenge (large number of spores) to the sterilization
process, so they are the only sterilization indicator
that provides a direct measure of lethality.

Biological indicators have evolved in consistency,

convenience, and speed (readout time).

Different technologies have been developed to

detect a earlier signals from the spores, allowing
faster readout times.

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Next Months 3M Sterile U Webinar

August 21, 2014:

Questions?

The Nuts and Bolts of Washer-Disinfectors

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