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Volume 24, Number 1—January 2018

Two-Center Evaluation of Disinfectant Efficacy against Ebola Virus in Clinical and Laboratory Matrices

Sophie J. Smither, Lin Eastaugh, Claire Marie Filone, Denise Freeburger, Artemas Herzog, M. Stephen Lever, David M. Miller, Dana Mitzel, James W. Noah, Mary S. Reddick-Elick, Amy Reese, Michael Schuit, Carly B. Wlazlowski, Michael Hevey, and Victoria Wahl-JensenComments to Author 
Author affiliations: Defence Science and Technology Laboratory, Porton Down, UK (S.J. Smither, L. Eastaugh, M.S. Lever); National Biodefense Analysis and Countermeasures Center, Frederick, Maryland, USA (C.M. Filone, D. Freeburger, D.M. Miller, D. Mitzel, J.W. Noah, M.S. Reddick-Elick, A. Reese, M. Schuit, C.B. Wlazlowski, M. Hevey, V. Wahl-Jensen); Censeo Consulting Group, Seattle, Washington, USA (A. Herzog)

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Table 1

Disinfection study parameters for evaluation of 6 disinfectants for reducing Ebola virus titers in dry blood or cell culture medium*

Parameter NBACC DSTL
Virus (dilution in blood)
Ebola virus/Makona-C05 (1:10)
Ebola virus/Makona-C07 (1:10)
Vero C1008, Vero 76, clone E6, Vero E6 
(ATCC CRL-1586)
Vero C1008, ECACC #85020206
Blood source
Human whole blood in EDTA 
(Bioreclamation IVT, Westbury, NY, USA)
Fresh whole blood in EDTA from male Porton rats
Blood droplet size (µL) and state
10, wet or dried
20, dried
Surface coupons
304 stainless steel (Diamond Perforated Metals, Visalia, CA, USA); 6061 Aluminum (Speedy Metals, New Berlin, WI, USA) used for experiments with dry medium (peracetic acid and acidified bleach) and wet blood (peracetic acid) only
3014 stainless steel
Sodium hypochlorite
Supplier Clorox (Oakland, CA, USA) Sychem (Leigh On Sea, UK)
Concentration tested, vol/vol 0.5%, 10% hypochlorite 1% hypochlorite
Volume tested, µL 30 or 100 100
Contact time, min 15 15
Peracetic acid
Supplier Sigma-Aldrich (St. Louis, MO, USA) Sigma-Aldrich
Concentration tested, vol/vol 5% 0.2%
Volume tested, µL 100 100
Contact time, min 5 10
Micro-Chem Plus
Supplier National Chemical Laboratories, Inc. (Winona, MN, USA) ND
Concentration tested, vol/vol 1.5%
Volume tested, µL 30 or 100
Contact time, min 10
Purell Advanced
Supplier GOJO Industries (Akron, OH, USA) ND
Concentration tested, vol/vol 70% ethanol
Volume tested, µL 30
Contact time, min 2
Steriplex SD
Supplier sBioMed LLC (Orem, UT, USA) ND
Concentration tested 0.015% silver, 10.000% ethanol, 0.020% H2O2, 0.150% peroxyacetic acid, 0.150% acetic acid, 0.075% inert food grade ingredients proprietary, 89.590% water
Volume tested, µL 100
Contact time, min 5
Acidified bleach
Supplier Clorox ND
Concentration tested, vol/vol 10% bleach (0.5% sodium hypochlorite) + 1% acetic acid
Volume tested, µL 100
Contact time, min
Neutralization 5 mL cell culture medium (10% FBS) or 
direct recovery using washing/filtration 2 mL cell culture medium (2% FBS)
Recovery method 30 μL bleach: none (5 mL medium only); 100 µL bleach; Steriplex SD: acidified bleach or peracetic acid; after neutralization in 5 mL medium, 2 mL was ultrafiltered in an Amicon (Bedford, MA, USA) 100-kDa NMWL unit, washed 3 times with 2 mL PBS after centrifugation at 5,000 × g for 10 min, and resuspended in 2 mL medium; Purell Advanced: diluted to 130 μL with PBS and recovered using a PD MultiTrap G-25 96-well microplate column array. Dilute supernatant to 5 mL with medium; Micro-Chem Plus: samples directly added to Amicon Ultra-0.5 (30- or 100-kDa NMWL) ultrafiltration unit, washed 3 times with 0.5 mL PBS by centrifugation at 5,000 × g for 10 min, and resuspended in 5 mL medium Washed by centrifugation 8,000 rpm for 5 min and resuspended in 1 mL medium

*Titers were assessed by using the 50% tissue culture infectious dose assay. ATCC, American Type Culture Collection; DSTL, Defence Science and Technology Laboratory; ECACC, European Collection of Authenticated Cell Cultures; FBS, fetal bovine serum; NBACC, National Biodefense Analysis and Countermeasures Center; ND, no data; NMWL, nominal molecular weight limit; PBS, phosphate-buffered saline.

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Page created: December 19, 2017
Page updated: December 19, 2017
Page reviewed: December 19, 2017
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