VirucidalPerformanceofVariousProfessionalHandHygieneProdu VirucidalPerformanceofVariousProfessionalHandHygieneProducts cts

AgainstAvianInfluenzaAH5N1 AgainstAvianInfluenzaAH5N1
CAMBondiMPH CAMBondiMPH
1 1
,JWArbogastPhD ,JWArbogastPhD
1 1
,DRMacingaPhD ,DRMacingaPhD
1 1
,RLambkin ,RLambkin WilliamsPhD WilliamsPhD
2 2
,EMoaneBBSc ,EMoaneBBSc
2 2
1.GOJOIndustries,Inc.,Akron,Ohio,USA2.RetroscreenVirologyLtd.,London,UK
ABSTRACT ABSTRACT
BACKGROUND/OBJECTIVES: BACKGROUND/OBJECTIVES: Influenza A is a pathogen of substantial public health concern worldwide. The evidence
for human-to-human transmission of Influenza A H5N1 has identified it as a strain with pandemic potential, which would
present significant challenges to the healthcare infrastructure in terms of both infection control and patient volume. While
standard hand hygiene practices are recommended by leading public health organizations like the CDC and WHO to
prevent transmission, quantitative data on the effectiveness of hand hygiene products against these strains are not
readily available. In an effort to determine the virucidal performance of hand hygiene products on the potential pandemic
strain Avian Influenza A H5N1, we evaluated the virucidal efficacy of various professional hand hygiene products
commonly used in healthcare settings against Avian Influenza A NIBRG-14 [H5N1] using in-vitro techniques.
METHODS: METHODS: 40L of virus was added to 360 L of test article and incubated at room temperature for 15 seconds. The
reaction was terminated by 10-fold (v/v) dilution in MDCK infection media, supernatant was titrated 10-fold (v/v) across a
96 well plate, and cells were incubated for 1 hour. After incubation, cells were washed twice with PBS, fresh infection
media was added and cells were incubated for 3 days. After the final incubation period, the Hemagglutination Assay (HA)
was used to determine the presence of virus.
RESULTS: RESULTS: Four currently marketed ethanol instant hand sanitizers (3 gel, 1 foam), an ethanol surgical hand antiseptic, a
Triclosan foaming handwash, and a Chloroxylenol foaming handwash exhibited complete kill (>3log to >4log reduction of
H5N1 virus above the detectable limit) after an exposure of only 15 seconds.
CONCLUSIONS: CONCLUSIONS: These data demonstrate that the well-formulated 0.3% foaming Triclosan and 0.5% Chloroxylenol hand
washes and the five 62%-81.4% alcohol-based hand antiseptics evaluated in this study are in fact capable of rapidly
inactivating Avian Influenza A H5N1. Although Influenza A is an enveloped virus, and thus generally considered
susceptible to various antimicrobial agents, these data provide scientific support for the CDC and WHO position on hand
hygiene as the primary prevention measure for Influenza infection in healthcare.
Avian Influenza A NIBRG-14 [H5N1] was procured from the
reference collection of the National Institute for Biological Standards
and Control, London, England. Cytotoxicity assays were performed
to determine the detection limit for each test substance. 40L of
virus was added to 360 L of test article and incubated at room
temperature for 15 seconds. The reaction was terminated by 10-fold
(v/v) dilution in MDCK infection media, supernatant was titrated 10-
fold (v/v) across a 96 well plate of MDCK cells and incubated for 1
hour. After incubation, cells were washed twice with PBS, fresh
infection media was added and cells were incubated for 3 days.
After the final incubation period, the Hemagglutination Assay (HA)
was used to determine the presence of virus, where the presence of
agglutination indicated virus presence. HA results were used to
calculate the virus titer using the Krber Calculation.
METHODS METHODS
Virucidal Assay Virucidal Assay CONCLUSIONS & DISCUSSION CONCLUSIONS & DISCUSSION
Our results show Our results show:
Alcohol-based instant hand sanitizers with active ingredient
levels 62% ethanol completely inactivate AIA H5N1 in 15 seconds
in-vitro.
Well-formulated handwashes with Triclosan (0.3%) or
Chloroxylenol (0.5%) are able to completely inactivate AIA H5N1 in
only 15 seconds in-vitro.
Avian Influenza can be effectively inactivated by hand hygiene
products that are currently being used in the healthcare setting.
Vigilant hand hygiene practices with efficacious hand hygiene
products should be a key component of Avian Influenza H5N1
infection control used in conjunction with respiratory precautions.
Until now the virucidal activity of commonly used hand hygiene
products versus H5N1 was unknown. The general opinion that
enveloped viruses are susceptible to and controllable with the
appropriate hand hygiene practices
1-2
has been confirmed in the case
of Avian Influenza A H5N1 by these results. Due to the perception of
Avian Influenza A H5N1 as a potential pandemic strain, there has
been an emphasis on airborne transmission over direct contact
transmission. As the natural transmission of AIA predominantly
occurs after direct contact, and AIA can survive on porous and non-
porous surfaces for 1-2 days and on hands for >5 minutes
5-6
, hand
hygiene has been identified by the World Health Organization as the
primary prevention measure for AIA H5N1 infection
2-4
. Therefore,
proper hand hygiene in the healthcare setting using products with
proven performance against AIA H5N1 is critical not only for
pandemic preparedness, but also as a key infection control
component for AIA H5N1 and other strains of influenza.
REFERENCES REFERENCES
1. Boyce JM, Pittet D. Guideline for Hand Hygiene in Health-Care Settings. Recommendations of the Healthcare Infection Control
Practices Advisory Committee and the HICPAC/SHEA/APIC/IDSA Hand Hygiene Task Force. Society for Healthcare Epidemiology of
America/Association for Professionals in Infection Control/Infectious Diseases Society of America. MMWR Recomm. Rep. 2002; 51(RR-
16):1-45.
2. World Health Organization. Prevention and control of influenza due to avian influenza virus A (H5N1). Version 30 January 2004.
3. Brankston G, Gitterman L, Hirji A, Lemieux C, Gardam M. Transmission of influenza A in human beings. Lancet Infect. Dis. 2007;7:257-
7.
4. Tellier R. Review of aerosol transmission of influenza A virus. Emerg. Infect. Dis. 2006;12(11):1657-62.
5. Kramer A, Schwebke I, Kampf G. How long do nosocomial pathogens persist on inanimate surfaces? A systematic review. BMC Infect.
Dis. 2006;6:130.
6. Inglesby TV, Nuzzo JB, OToole T, Henderson DA. Disease mitigation measures in the control of pandemic influenza. Biosecur.
Bioterror. 2006;4(4):376-83.
RESULTS RESULTS
Product Product
Active Active
Ingredient Ingredient
Exposure Exposure
Time Time
Log Log
10 10
Reduction Reduction
1 1
Percent Percent
Reduction Reduction
1 1
PURELL

Instant Hand Sanitizer 62% Ethanol 15 seconds >3.50 >99.96%

PURELL

Instant Hand Sanitizer with

Dermaglycerin System
62% Ethanol 15 seconds >3.79 >99.98%
GOJO

Medical Hand Sanitizer

(International Healthcare Product)
76.9% - 81.4%
Ethanol
15 seconds >4.50 >99.99%
PURELL

Instant Hand Sanitizer Foam 62% Ethanol 15 seconds >3.50 >99.96%

PURELL

Surgical Scrub 70% Ethanol 15 seconds >3.75 >99.98%

GOJO

PROVON

Foaming Handwash
with Triclosan and Moisturizers
2
0.3% Triclosan 15 seconds >4.00 >99.99%
GOJO

MICRELL

Antibacterial Foam
Handwash
0.5%
Chloroxylenol
15 seconds >4.25 >99.99%
40 40 L L H5N1 H5N1
360 360 L L Test Article Test Article
15 Second 15 Second
Incubation Incubation
Reaction Termination Reaction Termination
10 10- -fold Dilution ( fold Dilution (v/v v/v) )
MDCK Infection Media MDCK Infection Media
10 10- -fold Supernatant Titration & 1 fold Supernatant Titration & 1
Hour Incubation on MDCK Cells Hour Incubation on MDCK Cells
P
B
S

W
a
s
h

X
2
P
B
S

W
a
s
h

X
2
A
d
d

M
D
C
K

M
e
d
i
a
A
d
d

M
D
C
K

M
e
d
i
a
3 Day Incubation 3 Day Incubation
Hemagglutination Assay Hemagglutination Assay
for H5N1 Detection for H5N1 Detection
1. 1. A designation of A designation of > > denotes a complete inactivation of Avian Influenza A H5N1 denotes a complete inactivation of Avian Influenza A H5N1
2. To reduce 2. To reduce cytotoxicity cytotoxicity, the terminated reaction mixture was passed through a , the terminated reaction mixture was passed through a Sephadex Sephadex

column column
Cytotoxicity Cytotoxicity Assay Assay
4 hour 4 hour
incubation incubation
+ + tCPE tCPE
observation observation
PBS PBS
Wash x2 Wash x2
+ MDCK + MDCK
media media
added added
10 10- -fold fold
supernatant supernatant
titration + 1 titration + 1
hour hour
incubation incubation
Reaction Reaction
diluted diluted
10 10- -fold fold
in MDCK in MDCK
infection infection
media media
360 360 L L
test test
article + article +
40 40 L L
MDCK MDCK
infection infection
media media
K K rber rber Calculation Calculation
For additional information contact Cara Bondi, MPH: mccrackenc@gojo.com 2007 GOJO Industries, Inc., all rights reserved