Inactivation Methods to Prevent HCV

Inactivation methods to prevent HCV transmission among
people who inject drugs
PD Dr. Eike Steinmann
Centre for Experimental and Clinical Infection Research
Institute of Experimental Virology, Twincore*
*joint venture between Medical School Hannover and Helmholtz Centre for Infection Research
Stability of human pathogenic viruses
with envelope
strong lipophilic
character
without envelope
weak lipophilic
character
hydrophilic
character
lipophilic
character
Retroviridae
Poxviridae
Picornaviridae
Adenoviridae
Flaviviridae
Hepadnaviridae
Parvoviridae
Reoviridae
Togaviridae
Herpesviridae
Caliciviridae
E2
E1
Orthomyxoviridae
ss(+) RNA
nucleocapsid (core)
lipidmembrane
based on Klein and Deforest (1963)
Ways of HCV Transmissions
medical
procedures
drug abuse
sex 15%
i.v. drug abuse 60%
transfusions 10%
unknown
10%
transfusion
sex
vertical
other
Virucidal efficacy of different alcohols against HCV
Quantitative suspension:
100 µl virus
900 µl disinfectant
mix, incubation, mix
titration infectivity (TCID50/ml)
Effect of ethanol, 1-propanol and 2-propanol on HCV/BVDV
Log10 infectivity TCID50/ml
HCV
8
8
7
7
6
6
5
5
4
4
3
3
2
2
1
1
0
0
contr.
5%
10%
20%
30%
40%
ethanol
1-propanol
2-propanol
detection limit
contr.
5%
10%
20%
30%
40%
incubation time 5 min
incubation time 1 min
Log10 infectivity TCID50/ml
BVDV
8
8
7
7
6
6
5
5
4
4
3
3
2
2
1
1
0
ethanol
1-propanol
2-propanol
0
contr.
5%
10%
20%
incubation time 1 min
30%
40%
contr.
5%
10%
20%
30%
40%
incubation time 5 min
Ciesek et al. Journal of Infectious Diseases
HCV stability and inactivation in suspension
7
HCV+Serum
Log RLU/well
6
5
4
3
2
0
7
14
days
21
28
Log10 infectivity TCID50/ml
HCV
7
HCV
6
cytotoxicity
5
4
3
2
1
0
contr. 1
2
3
4
5
6
7
undiluted, 30 sec
Ciesek et al. Journal of Infectious Diseases 2010
Steinmann et al. American Journal of Infection Control 2010
Steinmann et al. Antimicrobial Resistance and Infection
Establishment of a HCV carrier assay
virusinoculum
50 µL
(drying)
Log10 Infectivity TCID50/ml
Log10 Infectivity TCID50/ml
900 µL
medium Ø FCS
(1 min vortex)
(incubation for 1 min)
titration infectivity
5
7
6
5
4
3
2
1
0
100 µL
test substance
suspension
dried
4
3
2
1
0
Control 10
20
30
40
50
1-propanol
60
10
20
30
40
50
2-propanol
60
10
20
30
40
50
60%
ethanol
Dörrbecker et al. Journal of Infectious Diseases
Survival of dried HCV on inanimate surfaces
Log10 Infectivity TCID50/ml
6
HCV
cytotoxicity
5
4
3
2
1
0
Control 0.025 0.25
0.50 0.025 0.25
C
B
A
5
Log10 Infectivity TCID50/ml
0.50 0.025 0.25
HCV - serum
HCV + serum
4
3
2
1
0
0
1
2
3
4
days
5
6
7
8
0.50 0.025 0.25
D
0.50%
Development of a drug transmission assay
5
Log10 RLU/well
4
3
2
1
0
15
25
35
45
55
65
temperature in °C
75
Dörrbecker et al. Journal of Infectious Diseases
Transmission of Hepatitis C virus among injecting drug users:
viral stability and association with drug preparation equipment
4
500 µl
3
200 µl
40 µl
2
8 µl
1.5 µl
1
0
0
1
2
4
6
8
10 12
days
14
16
18
20
5
virus spike
Log10 Infectivity TCID50/ml
Log10 Infectivity TCID50/ml
5
recovery concentrated
4
3
2
1
0
plastic
aluminium
glass
container material
Dörrbecker et al. Journal of Infectious Diseases
Transmission of Hepatitis C virus among injecting drug users:
viral stability and association with drug preparation equipment
Dörrbecker et al. Journal of Infectious Diseases
Thermo-stability of seven Hepatitis C virus genotypes in vitro
and in vivo
1b
2a
3a
4a
5a
6a
7a
9
8
7
6
5
4
3
2
1
0
0
7
14
21
28
35
time [days]
Log10 HCV RNA [IU/mL]
Log10 Infectivity [TCID50/ml]
1a
10
1a
1b
2a
3a
4a
5a
6a
7a
pool
Dörrbecker et al. Journal of Viral Hepatitis 2
Inactivation of HCV and HIV by microwave!?
Inactivation of HCV and HIV by microwave
HCV
HCV/HIV
HIV
HIV/HCV
Inactivation of HCV and HIV by microwave
Inactivation of HCV and HIV by microwave
Inactivation of HCV and HIV by microwave!?
Acknowledgement
Twincore, Institute of Experimental Virology
Thomas Pietschmann
Dorothea Bankwitz
Pia Banse
Patrick Behrendt
Richard Brown
Janina Brüning
Wladislaw Butovetsky
Stephanie Pfänder Stephanie Walter
Juliane Dörrbecker
Anggakusuma
Aci Siddharta
Mandy Döpke
Svenja Drave
Michael Engelmann
Cooperations
Gisa Gerold
Sabine Giese
S. Ciesek, T. v. Hahn, H. Wedemeyer, M.P. Manns
Corinne Ginkel
P. Meuleman
Ghent, Belgium
J. Cavalleri, K. Feige
Hannover, Germany
Sibylle Haid
W. Baumgärtner, K. Hahn
Hannover, Germany
Kathrin Hüging
P. Becher, A. Postel
Hannover, Germany
Sebastian Joecks
M. Heim, B. Campana
Basel, Switzerland
Christina Grethe
Alex Hahn
Tanvi Khera
Paula Perin
Julie Sheldon
Daniel Todt
Gabrielle Vieyres
Romy Weller
Kathrin Welsch
AG Goffinet
Funding:
Hannover, Germany