Meathods for the Evaluation of Hand Disinfectants
Dr. Manfred Rotter, University of Vienna, Austria
Sponsored by Deb Hand Hygiene [Link]
Methods for the Evaluation of
Hand Disinfectants
Evaluation of Hand Disinfectants (1)
Parameter: Reduction of bioburden
In vitro
MIC of disinfectant vs. selected strains
MBC of disinfectant vs. selected strains
Quantitative suspension tests for
(bacteri-, fungi-, viru-)cidal properties
Kill-time studies suspension tests
Manfred L. Rotter
Institute of Hygiene and Medical Microbiology
Medical University of Vienna
Sponsored by
Hosted by Paul Webber
paul@[Link]
[Link]
[Link]
Rotter/teleclass/2005/Bo
Rotter/teleclass/2005/Bo
Evaluation of Hand Disinfectants (3)
Parameter: Reduction of infections
Evaluation of Hand Disinfectants (2)
In vivo
Controlled laboratory tests simulating
practical conditions on hands of volunteers
Clinical trial
Comparative trial with the aim
Novum > Reference
Field trials
Equivalence study
Rotter/teleclass/2005/Bo
Rotter/teleclass/2005/Bo
SAMPLE SIZE NECESSARY FOR SIGNIFICANT
DIFFERENCE OF PROPORTIONS
(Example for comparative trial)
PRESENT RATIO OF HAND-TRANSMITTED
NOSOCOMIAL INFECTIONS be
Desired reduction of infection ratio be
50 %
NEW RATIO OF HAND-TRANSMITTED
NOSOCOMIAL INFECTIONS intended be
Level of significance (one-sided)
Power of statistical test
Laboratory Test Methods
2%
1%
Hygienic Hand Disinfection:
= 5 %
1- = 90%
SAMPLE SIZE
(= Number patients per experimental arm)
(Hygienic Hand Wash and Hygienic Hand Rub)
In vitro
In vivo
2500
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Meathods for the Evaluation of Hand Disinfectants
Dr. Manfred Rotter, University of Vienna, Austria
Sponsored by Deb Hand Hygiene [Link]
Bactericidal Properties of Hand Disinfectants
Proposed Suspension test prEN 12054
In vitro
Test organisms
Staphylococcus aureus
Pseudomonas aeruginosa
Escherichia coli K 12
Enterococcus hirae
20C
Temperature
Contact time
- hygienic
- surgical
Requirement
1 min (optional 0,5 min)
5 min (optional 1,2,3,4 min)
reduction: rub 5.0 lg,
wash 3.0 lg
7
Rotter/teleclass/2005/Bo
Hygienic Handwash with antiseptic soap
EN 1499. Simulating practical Conditions
In vivo
Volunteers
Test organism
Recovery
Application
- product
- reference
Requirement
Discrimination
Mean log reduction factor (log 10)
Hygienic Handrub EN 1500
Simulating practical Conditions
In vivo
Volunteers
Test organism
Recovery
Application:
- product
- reference
12-15
Escherichia coli K 12
Fingertip rub before and after treatment
30 or 60 s, according to manufacturer
60 s handwash with unmedicated soap
Product significantly (p = 0.01 unidirectional)
more efficacious than soap
Means > 0.5 lg different (Power: 0.90)
Rotterr/teleclass/2005/Bo
Requirement
Discrimination
12-15
Escherichia coli K 12
Fingertip rub before and after treatment
30 or 60 s, according to manufacturer
2 x 30s (=60 s) handrub with 2x3 ml
60% (vol) 2-propanol
Product not significantly (p = 0.1, unidir.)
less efficacious than 2-propanol 60%, 1min
Means > 0.6 lg different (Power: 0.95)
Rotter/CHICA/Desinf. 03
Rotter/teleclass/2005Bo
10
Hygienic Hand Disinfection ANOVA: non-standardized
Results (lg RFPi): 5 agents, 5 repetitions, 2 laboratories,
with 15 volunteers in each
5,0
4,0
3,0
Place . . .
VM
V M B
V M B
V M B
Preparation: n-Propanol Iso-Propa- Ethanol 70% Povidone-I50%
nol 60%
Standard
+ chlorhex.
0,5 %
0,75% soap
Source of
Variation
DF
Mean
Squares
Agents (5)
Volunteers (15)
Laboratories (2)
4
14
1
102,4
2,6
5,8
327,5
8,1
18,6
<0,0001
<0,0001
<0,0001
PxV
42
0,7
2,2
<0,0001
VxL
3,4
10,8
<0,0001
PxL
14
3,1
10,0
<0,0001
PxVxL
42
0,7
2,4
<0,0001
470
0,3
V M B
Phenolic soap
Error
11
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Page 2
Meathods for the Evaluation of Hand Disinfectants
Dr. Manfred Rotter, University of Vienna, Austria
Sponsored by Deb Hand Hygiene [Link]
Health-Care Antiseptic Drug Products-FDA (1)
Hygienic Hand Disinfection ANOVA:
standardized results (lg RFPi,-lg RFRi):
4 [productsreference], 5 repetitions , 2 laboratories,
with 15 volunteers in each
Source of
Variation
Products-Reference (4)
DF
3
Mean
SQ
90,2
Volunteers (15)
14
2,7
Laboratories (2)
In vitro
Antibacterial spectrum of
117,5
<0,00001
3,5
<0,00001
3,5
4,6
n.s.
PxV
42
0,9
1,1
n.s.
VxL
1,6
2,1
n.s.
PxL
14
1,7
2,1
<0,01
PxVxL
42
0,9
1,2
n.s.
466
0,8
Error
Rotter/WHO/2004/Bo
- active ingredient
- carrier
- both
MICs with 20 microbial species, 50 strains of each
(50% fresh clinical strains)
- 9 gramnegative spp.
-10 grampositive spp.
- Candida (incl. C. albicans)
(= approx. 1000 strains)
Development of resistance study
Time-kill study (suspension test) with selected strains for
0, 3, 6, 9, 12, 15, 20, 30 min
Rotter/teleclass/2005/Bo
13
14
Standard Test Method for Determining the
Bacteria
- eliminating Effectiveness of Hygienic
Handwash and Handrub.
ASTM E
- 2276
Health-Care Antiseptic Drug Products - FDA (2)
Antiseptic handwash and Health-Care Personnel handwash
(originally ASTM -E 1174)
In vivo
In vivo
Volunteers (N):
approx. total of 108
- Test preparation:
approx. 54
- Positive control:
approx. 54
Contaminant bacterium:
Serratia marcescens , (E. coli)
Contamination and application: 10 times on an experimental day
(Test and control in parallel)
Samplings:
- after 1st contamination (baseline)
- after 1st, 3rd, 7th, 10th wash (rub)
Required reduction within 5min: - after 1st handwash:
2 lg
- after 10th handwash: 3 lg
Rotter/teleclass/2005/Bo
(Similar to Virus-eliminating Test acc. to ASTM E-1838)
Finger Pads including thumbs of at least 2 volunteers
Test bacteria: S. marcescens, E. coli, S. aureus, [Link].
Requirement: not defined, but in comparison to a
negative and positive control
15
16
Virucidal Hand Disinfection Test ASTM E-1838
Preparation for the test
Virucidal Hand DisinfectionTests
Suspension Tests
prEN 14476
DVV
Polio 1
Adeno 5
Polio 1
Adeno
Vaccinia
4.0 lg
SV 40
4.0 lg
In vivo Tests
Marking of test areas
Finger pad
Whole hand
ASTM E-1838
ASTM E-2011
Contamination with 10 l Virus suspension
(acc. to Sattar)
( Steinmann)
Adeno (human 4)
Rota (human) Wa Rota (human)
Rhino (human) 37 Rhino (human)
Hepatitis A HM-175
not defined
not defined
Test area with wet
Virus suspension
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Test area with dried
Virus suspension
2030 minutes
18
Page 3
Meathods for the Evaluation of Hand Disinfectants
Dr. Manfred Rotter, University of Vienna, Austria
Sponsored by Deb Hand Hygiene [Link]
Virucidal Hand Disinfection Test ASTM E-1838
Wet virus suspension
dried virus suspension
10s Exposition vs 1ml of
Water
Product
Laboratory Test Methods
Rinse with 15 ml WSH for 5 s
Surgical Hand Disinfection:
Virus elution in 1 ml water of standardized
hardness during 10 20 s (40 Inversions)
(Surgical Hand Wash and Surgical Hand Rub)
scrape off FP on rim
In vitro
In vivo
Virus count
19
Laboratory Test Methods
Laboratory Test Methods
Surgical Hand Disinfection:
In vitro
Surgical Hand Disinfection:
In vivo
Same Test Method as for
Hygienic Hand Disinfection
Surgical Handrub/wash prEN 12791
Simulating practical conditions
In vivo
Volunteers (N):
18-20
Test organism:
Resident skin flora
Recovery
Fingertip rub before and after treatment
Application of
- Product:
acc. to manufacturer, max.5 min handrub/wash
- Reference:
3 min handrub with 60%/vol 1-propanol (nx3 ml)
Immediate effect sample: immediately after end of treatment (one hand)
3-hours effect sample:
3 hrs after end of treatment (other [gloved] hand)
Requirement:
Product not significantly (imm: p=0.1; 3-hrs: 2p =0.01)
less efficacious than reference
Discrimination:
imm. Effect: Means > 0.5 lg different (Power: 0.95)
Sustained effect:
Optional claim: At 3 hrs, product significantly (p=0.01,
unidirectional) more efficacious than reference
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Page 4
Meathods for the Evaluation of Hand Disinfectants
Dr. Manfred Rotter, University of Vienna, Austria
Sponsored by Deb Hand Hygiene [Link]
Surgical Hand Disinfection - ANOVA :
non-standardized results (lgRFi):,
4 agents , 2 hands, 5 laboratories, 20 volunteers in each
Surgical Hand Disinfection according to EN 12791
4 disinfectants, 5 laboratories, both hands
Mean (N=20) lg reductio
3,5
3,0
2,5
2,0
1,5
1,0
0,5
0,0
Hands
Agents
Conc.
l
r
n-propanol
60%
l
r
ethanol
85%
l
r
iso-propanol
70%
r
CHG
4%
25
Source of
Variation
1 Agents (4)
2 Laboratories (5)
3 Hands (2)
DF
1x2
1x3
2x3
1x2x3
53,97
< 0,001 *
5,22
< 0,001 *
4,93
0,029
12
4
3
12
1,67
1,37
0,66
1,55
0,073
0,249
0,579
0,107
* significant at p 0,01
Rotter/WHO 2004/Bo2003
26
Surgical Hand Disinfection - ANOVA :
standardized results (lgRFPi - lgRFRi):
3 [products reference], 5 laboratories, 2 hands,
20 volunteers in each
Source of
Variation
1 [Products-Reference] (3)
2 Laboratories (5)
3 Hands (2)
1x2
1x3
2x3
1x2x3
DF
42,16
1,67
0,162
0,70
0,404
8
4
2
8
1,66
3,41
0,63
0,40
0,109
0,012
0,533
0,918
* significant at p 0,01
< 0,001 *
Rotter/WHO 2004/Bo2003
27
Surgical Hand Disinfection: FDA (orig. ASTM -E 1115)
REDUCTION OF RELEASE OF SKIN BACTERIA
EFFICACY OF SURGICAL HAND DISINFECTION (5 min)
Volunteers (N):
approx: 100 (150)
per arm:
- Test:
approx. 50
- Positive control: approx. 50
- ( Placebo )
( 50 )
LIQUID SOAP
0.0 0.0
POVIDONE-IODINE
LIQUID SOAP 0,5 %
CHLORHEXIDINEDETERGENT 4 %
POVIDONE-IODINE
AQU. SOLUTION 10 %
ISO-PROPANOL 60 %
HEXACHLOROPHENEDETERGENT 3 %
ISO-PROPANOL 70 %
90.0 1.0
99.0 2.0
Testbacteria:
normal resident handflora
Application of Product:
acc. to manufacturers instruction or
without any: apply product 2 x 5 min, then
rinse hands for 1 min
Baseline:
rinse hands for 30 s, wash hands for 30s,
rinse hands for 30s
Positive control:
FDA-approved antiseptic; all parameters
as product; concurrent testing
N-PROPANOL 60 %
ISO-PROPANOL 70% +
CHLORHEXIDINE 0,5%
99.9 3.0
60
28
180 min
AFTER DISINFECTION
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Meathods for the Evaluation of Hand Disinfectants
Dr. Manfred Rotter, University of Vienna, Austria
Sponsored by Deb Hand Hygiene [Link]
Surgical Hand Disinfection: FDA in-vivo model
Schedule for disinfecting and sampling and required
lg bacterial reduction
Sampling
Times (hrs)
Day of test periode
1 2 3 4 5
1lg
x
O x
O 2lg
x
<bl
1/60
3
6
x
x
x
O
O
3lg
X: Desinfection: Day 1 (1/60), day 2, 3, 4 (1/60, 3, 6), day 5 (1/60)
O: Sampling: Day 1, 2 and 5: after 1/60, 3 and 6 hrs with gloves
Further Information is available!
bl: Baseline
Rotter/teleclass/2005
31
Other 2005 Teleclasses
For more information, refer to
[Link]/[Link]
April 21 CreutzfeldtCreutzfeldt-Jakob Disease: Recommendations for Disinfection
April 28
and Sterilization with Dr. William Rutala
Overcoming the Resistance of Biofilms with Dr. Peter Gilbert
Sponsored by Virox Technologies Inc. [Link]
May 19
Antiseptic Practice & Procedure with Susan Crow
May 26
Canadian Response to West Nile Virus with Dr. Paul Sockett
June 7 Measuring the Cost of Hospital Infection with Dr. Barry Cookson
Sponsored by 3M Canada [Link]
Questions?
Contact Paul Webber paul@[Link]
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