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3M Infection Prevention Solutions

3M™ Sterile U Web Meeting – July 17, 2014 The Science of Speed – The Evolution of Biological Indicators

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Sterilization Monitoring - Quality Control

 
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Load Control Pack Control Equipment Exposure Control Control
Load
Control
Pack
Control
Equipment
Exposure
Control
Control

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Welcome! Facilitators: Christophe de Campeau, 3M Questions Mute feature (*7 = unmute; *6 = mute)
Welcome!
Facilitators:
Christophe de Campeau, 3M
Questions
Mute feature (*7 = unmute; *6 = mute)
“Chat” feature
Technical difficulties
Post session follow-up
For more information: www.3M.com/3MSterileU
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Topic: The Science of Speed – The Evolution of Biological Indicators

Sandra Velte, 3M

– The Evolution of Biological Indicators Sandra Velte, 3M Speakers: Craig Wallace – Senior Technical Specialist

Speakers: Craig Wallace – Senior Technical Specialist – 3M Housekeeping

Craig Wallace – Senior Technical Specialist – 3M Housekeeping Craig Wallace – 3M © 3M 2014.

Craig Wallace – 3M

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Learning Objectives

1. 2. 3. 4
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2.
3.
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Describe the design and function of biological indicators

Discuss how biological indicator incubation time is determined

Understand how rapid readout biological indicator systems work

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Sterilization Process Monitoring Tools

Biological Chemical Physical Indicators Indicators Monitors © 3M 2014. All Rights Reserved. 6
Biological
Chemical
Physical
Indicators
Indicators
Monitors
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The history of biological indicators

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First biological indicators

Robert Koch – 1881 Inoculated carriers © 3M 2014. All Rights Reserved. 9
Robert Koch – 1881
Inoculated carriers
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BI Basics

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Late 1800s

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Late 1800s 8 © 3M 2014. All Rights Reserved.

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National Institutes of Health (NIH) – 1955

Recommends Bacillus stearothermophilus* as the test organism for steam sterilization Forms spores - challenges process
Recommends Bacillus stearothermophilus* as the test organism for
steam sterilization
Forms
spores - challenges process
No threat of pathogenicity, pyrogenicity, or toxicity
* Now known as
Geobacillus
stearothermophilus
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The basic principle of biological indicators

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Direct measurement of lethality

Large 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

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

from a hostile environment, such as lack of nutrients © 3M 2014. All Rights Reserved. e.g.

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e.g. some Bacillus sp., Clostridium sp.

Protective shell around DNA, almost no water

No biological activity - dormant

Google Images
Google Images

Bacterial Spores - Formation

Google Images © 3M 2014. All Rights Reserved. 15
Google Images
© 3M 2014. All Rights Reserved.
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BacterialBacterial GrowthGrowth

“Bacterial Growth”

If we had a starting count of 10 contaminating bacteria that can grow and double every 20 minutes, you will have:

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Elapsed Time

# of Bacteria

0:00 10 0:20 20 0:40 40 1:00 80 1:20 180 1:40 360 2:00 720 2:20
0:00
10
0:20
20
0:40
40
1:00
80
1:20
180
1:40
360
2:00
720
2:20
1,440
2:40
2,880
3:00
5,760
3:20
11,520
3:40
23,040
4:00
46,080
4:20
92,160
4:40
184,320
5:00
368,640
5:20
737,280
5:40
1,474,560
6:00
2,949,120
Cycle time:
20 minutes
- 3 hours
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Bacterial Spores

Geobacillus stearothermophilus 14
Geobacillus
stearothermophilus
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Bacterial Spores Geobacillus stearothermophilus 14 Bacillus atropheus © 3M 2014. All Rights Reserved.

Bacillus

atropheus

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Bacterial Spores – Activation and germination

Many complex steps – driven by enzymes Google Images
Many complex steps – driven by enzymes
Google Images

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Types of BIs – Inoculated devices

 
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© 3M 2014. All Rights Reserved. Types of BIs – Inoculated devices   18   ©
© 3M 2014. All Rights Reserved. Types of BIs – Inoculated devices   18   ©
 

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Types of BIs – Spore Strips

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Types of BIs – Self-contained (SCBI)

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Filter Spore Strip Cap Growth Sleeve Media with pH Indicator
Filter
Spore
Strip
Cap
Growth
Sleeve
Media
with pH
Indicator

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Biological indicator performance requirements – ANSI/AAMI/ISO 11138 series © 3M 2014. All Rights Reserved. 23
Biological
indicator
performance
requirements –
ANSI/AAMI/ISO
11138 series
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Types of BIs – Spore Ampoules

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Self-contained BIs – color change

Growth of the test organisms produces acid pH of the growth media drops pH indicator
Growth of the test organisms produces acid
pH of the growth media drops
pH indicator changes color – positive BI
Negative
Positive
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Biological indicator testing - resistometer

© 3M 2014. All Rights Reserved. 24
© 3M 2014. All Rights Reserved.
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Key performance parameters

Population (number of test organisms)

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Number 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. 10 6 = 1,000,000)

D-value (resistance)

A measure of how resistant the organisms are to the process (how hard to kill) Measured as the time to reduce the population by 90% Expressed in time units (usually minutes)

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The evolution of BI technology © 3M 2014. All Rights Reserved. 27
The evolution of BI technology
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BI evolution

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Consistency

Purified cultures of carefully selected test organisms

Convenience

Self-contained designs eliminate transfers to growth media

Readout time

7 days reduced to minutes

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Biological indicators – key specifications

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Moist heat

D-value: 1.5 minute minimum (121°C)

Population: 1 x 10 5 CFU (minimum)

ISO 11138:2006
ISO 11138:2006

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Ethylene oxide

Ethylene oxide D-value: 2.5 minutes minimum Population: 1 x 10 6 CFU minimum

D-value:

2.5 minutes minimum

Population: 1 x 10 6 CFU minimum

Ethylene oxide D-value: 2.5 minutes minimum Population: 1 x 10 6 CFU minimum
Ethylene oxide D-value: 2.5 minutes minimum Population: 1 x 10 6 CFU minimum
Ethylene oxide D-value: 2.5 minutes minimum Population: 1 x 10 6 CFU minimum
Ethylene oxide D-value: 2.5 minutes minimum Population: 1 x 10 6 CFU minimum
© 3M 2014. All Rights Reserved. 28

© 3M 2014. All Rights Reserved.

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Biological indicator readout time

Readout (or incubation) time: Time required to decide that a biological indicator is NEGATIVE 7
Readout (or incubation) time: Time required to
decide that a biological indicator is NEGATIVE
7 days is the accepted readout time for BIs for
established sterilization processes, with known
test organisms
ISO/ANSI/AAMI 11138-1: 2006
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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.

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to an appropriate level of accuracy as compared to the 7 day result. 31 How is

How is that done?

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Evolution of readout times

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?????

Evolution of readout times 33 ????? 7 days 24 - 48 hours 1-4 hours 30 -

7 days

Evolution of readout times 33 ????? 7 days 24 - 48 hours 1-4 hours 30 -
Evolution of readout times 33 ????? 7 days 24 - 48 hours 1-4 hours 30 -

24 - 48 hours

1-4 hours

30 - 60 minutes

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BacterialBacterial GrowthGrowth

“Bacterial Growth”

If we had a starting count of 10 contaminating bacteria that can grow and double every 20 minutes, you will have:

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Elapsed Time

# of Bacteria

0:00

10

0:20

20

0:40

40

1:00

80

1:20

180

1:40

360

2:00

720

2:20

1,440

2:40

2,880

3:00

5,760

3:20

11,520

3:40

23,040

4:00

46,080

4:20

92,160

4:40

184,320

5:00

368,640

5:20

737,280

5:40

1,474,560

6:00

2,949,120

Cycle time: 20 minutes - 3 hours
Cycle time:
20 minutes
- 3 hours

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FDA Guidance – Reduced Incubation Time “standard seven or more days” “more than 97% “
FDA Guidance –
Reduced
Incubation Time
“standard seven or
more days”
“more than 97%
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Rapid biological indicator technologies*

*Less than 24 hours 34
*Less than 24 hours
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Mesa Labs Smart-Read™ Technology

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Standard SCBI growth media color change

Colorimeter reads color shift and declares a positive when certain color change criteria are met

a positive when certain color change criteria are met www.mesalabs.com © 3M 2014. All Rights Reserved.

www.mesalabs.com

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Readout time is 10 hours

Smart-Read™ is a registered trademark of Mesa Labs, Inc.

Bacterial spores – activation and germination

Many complex steps – driven by enzymes Google Images © 3M 2014. All Rights Reserved.
Many complex steps – driven by enzymes
Google Images
© 3M 2014. All Rights Reserved.
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Spore start up

Computer start up

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Spore start up Computer start up 38 © 3M 2014. All Rights Reserved.

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3M Rapid Attest and Super Rapid Attest SCBIs Technology: 3M Rapid Attest™ and Super Rapid
3M Rapid Attest and Super Rapid Attest SCBIs
Technology: 3M Rapid Attest™ and
Super Rapid Attest ™ Technology
Detects biological activity very early in the
spore’s activation and germination process
Growth
Media with
Super Rapid Attest
Takes advantage of the activity of enzymes used by the cell
to germinate and grow
pH Indicator
Spore
Cap
Sleeve
and
carrier
fluorescent
Detects the end product of the enzyme reactions
indicator
Glucosidase enzymes - carbohydrate metabolism
Geobacillus stearothermophilus: alpha-glucosidase
Bacillus atrophaeus: beta-glucosidase
Rapid Attest
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© 3M 2014. All Rights Reserved.
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3M Rapid Readout Technology Non-fluorescent substrate in growth media Fluorescent by-product (4-MU) detected in
3M Rapid Readout Technology
Non-fluorescent substrate
in growth media
Fluorescent by-product (4-MU)
detected in Auto-reader
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3M Rapid Readout Technology Non-fluorescent substrate in growth media Fluorescent by-product (4-MU) detected in
3M Rapid Readout Technology
Non-fluorescent substrate
in growth media
Fluorescent by-product (4-MU)
detected in Auto-reader
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Readout Times

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Rapid Attest 1292

Super Rapid Attest 1492V

3 hours

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.

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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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Questions?

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Rapid Attest and Super Rapid Attest also can provide a conventional outgrowth- based pH color
Rapid Attest and Super Rapid Attest also
can provide a conventional outgrowth-
based pH color change response
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Next Month’s 3M™ Sterile U Webinar

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August 21, 2014:

“The Nuts and Bolts of Washer-Disinfectors”

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