Volume 24, Number 1—January 2018
Dispatch
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-Jensen
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) |
| Cells |
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 |
| Disinfectant | ||
| 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 |
15 |
|
| Method | ||
| 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.
Page created: December 19, 2017
Page updated: December 19, 2017
Page reviewed: December 19, 2017
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