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Background: The use of disinfectant wipes in hospitals is increasing over the last years. These wipes should be able to inactivate microorganisms including viruses on environmental surfaces and to prevent their transfer to clean areas. The European norm (EN) 16615:2015 describes a wiping process over four fields starting on the contaminated field 1 followed by fields 2–4 and back to the starting point (4-field test). This test method exclusively describes killing and transfer of vegetative bacteria and fungi by disinfectant wipes without measuring virucidal activities. Therefore, it was the aim of this study to use the existing test methodology additionally to evaluate virus inactivation by wipes. Methods: The 4-field test was performed with four commercially available disinfectant wipes including the examination of the active solutions of these wipes with a reference wipe. Murine norovirus (MNV) as surrogate of human noroviruses, adenovirus (AdV) type 5 and polyomavirus SV40 (SV40) were chosen as test viruses. Results: The per acetic acid (PAA)-based wipe (wipe A) was able to inactivate all three test viruses resulting in a four log reduction on test field 1, whereas the quaternary ammonium compound (QAC)-based products (wipes B and C) failed to reach such reduction. Both QAC-based wipes were able to inactivate SV40 and only the active solution of wipe B was effective against MNV. Another wipe with 2-propanol as active ingredient (wipe D) was not able to show a sufficient efficacy against all three test viruses. There was a good agreement between the results of the wipes and the corresponding fluids showing no influence of the material of wipes. Tests with the 2-propanol-based wipe D showed a transfer of all test viruses to the non-contaminated test fields 2–4. SV40 was additionally transferred by the QAC-based wipe C with 0.78% active ingredients to these additional fields. In all other cases no virus transfer to test fields 2–4 was observed. Finally, no virus could be detected in the PAA-based wipe A after usage in the 4-field test in contrast to the other wipes examined. Conclusions: The successful performance of a 4-field test with viruses demonstrated that the existing wiping method with bacteria and fungi can be used in addition for measuring virucidal efficacy. The virus-inactivating properties of surface disinfectants could be evaluated therefore with a test simulating practical conditions with mechanical action resulting in more reliable data than the existing quantitative suspension tests and/or a carrier test without any mechanical action. Keywords: Virucidal efficacy, Wipes, 4-field test, Virus transfer, Surfaces * Correspondence: email@example.com Dr. Brill + Partner GmbH Institute for Hygiene and Microbiology, Norderoog 2, 28259 Bremen, Germany Full list of author information is available at the end of the article © The Author(s). 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Becker et al. Antimicrobial Resistance and Infection Control (2019) 8:121 Page 2 of 8 Background EN 16615:2015 . Here, the ability of disinfectant Microorganisms like gram-positive and gram-negative wipes to remove bacteria and fungi from a contaminated bacteria can persist on inanimate surfaces for prolonged test field 1 and the potential transfer between clean sur- time . In addition, Kramer and co-workers described faces (test fields 2–4) can be measured. Until now, this that viruses can persist for few hours up to months . method was only described with different bacteria and Furthermore, it was noted that in general the group of Candida albicans. In parallel, a test of hygienic towelette non-enveloped viruses are more stable on environmental wipe efficiency was developed for bacteria with the surfaces than the enveloped ones . Wiperator (Filtaflex ltd, Almonte, Ontario, Canada, Surfaces can become contaminated by hands, objects, http://www.filtaflex.ca/wiperator.htm), which is now the settling of virus containing aerosols or contaminated basis of the ASTM E2967–15 . Meanwhile, this fluids . Therefore, these surfaces may play an import- ASTM method was evaluated carefully in three inde- ant role for transmission of pathogens in the hospital [3, pendent laboratories . Here, two species of vegetative 4]. In contrast, the detection of viruses on environmen- bacteria, one gram-positive coccus (Staphylococcus aur- tal surfaces in the hospitals is rarely reported. Gallimore eus) and one gram-negative bacillus (Acinetobacter bau- et al. described the detection of mainly norovirus and mannii) were chosen as microorganisms but again no rotavirus in environmental swabs from two pediatric viruses. In addition, data with both vegetative bacteria wards . In an adult intensive care unit in Brazil group and Clostridioides difficile spores have been recently A rotavirus (RV-A) was detected by reverse-transcriptase published using detergent wipes . polymerase chain reaction (PCR) in environmental sur- Importantly, until now no data with viruses have been face samples. Here RV-A was regarded as biomarkers for generated with both methods described above. There- viral contamination . In addition, astroviruses were fore, it is still unclear whether the existing 4-field test found in a pediatric primary immunodeficiency unit . method or the Wiperator technology can be transferred In a pediatric outpatient waiting area most frequently to test methods using viruses. Finally, an appropriate adenovirus was detected in environmental samples . virus-inactivating claim should be possible and should In summary, it was estimated that more than 30% of all help to prevent nosocomial virus infections. hospital-acquired infections in many pediatric settings We used the European test method described for bac- may be caused by viruses . teria EN 16615:2015  by incorporating important non- Meanwhile, contaminated surfaces like “high-touch” enveloped viruses and started to develop a test for wipes environmental surfaces (HITES) in critical areas of the measuring inactivation and transfer to clean areas in one hospitals were identified  and these HITES may be manual step. The choice of viruses was influenced by the also responsible as vehicles for human pathogenic vi- existing test viruses from in vitro tests like the EN 14476: ruses. Therefore, careful cleaning and disinfection of en- 2015  and the German Guideline of Deutsche Vereini- vironmental surfaces in hospitals and medical wards is gung zur Bekämpfung der Viruskrankheiten e.V. (DVV) an important step in infection control and part of many and Robert Koch-Institute (RKI) . prevention programs in healthcare. The methods are based on a wide range of technologies including liquid Materials and methods disinfectants, self-disinfecting surfaces like copper or sil- Test viruses ver and the vaporisation of peroxides and other chemi- The murine norovirus S99 (MNV) was obtained from cals . Dr. E. Schreier, former head of FG15 Molecular Epi- Disinfection of surfaces by manually performed wiping demiology of Viral Pathogens at the RKI in D-13302 as one important part in the healthcare setting increased Berlin. The adenovirus (AdV) type 5 strain adenoid 75 in the last years. Recently, it has been shown that a pre- originated from PD Dr. A. Heim, Institute of Medical impregnated wipe with sporicidal efficacy demonstrated Virology, Hannover Medical School, D-30625 Hannover superiority in comparison to a cloth soaked in 1,000 and the polyomavirus SV40 strain 777 (SV40) was ob- ppm chlorine solution underlining the increasing im- tained from PD Dr. A. Sauerbrei, Institute of Virology portance of disinfectant wipes . and Antiviral Chemotherapy at the Friedrich Schiller For disinfectant wipes, the virucidal testing often starts University of Jena, D-07747 Jena. with a quantitative suspension test with the soaking so- lution or the squeezed-out liquid followed by a test Virus propagation simulating practical conditions. However, there is no The test virus suspensions were prepared by infecting European Norm (EN) measuring the virus-inactivating monolayers of the respective cell lines. The virus titres of 6 9 properties by wiping. these suspensions ranged from 10 to 10 TCID /ml. After intensive work by the group of J. Gebel in Bonn, MNV was propagated in RAW 264.7 cells (a macrophage- Germany a “4-field test” was developed which is now the like, Abelson leukemia virus transformed cell line derived Becker et al. Antimicrobial Resistance and Infection Control (2019) 8:121 Page 3 of 8 from BALB/c mice, ATCC TIB-71) and adenovirus in A549 measuring of efficacy of disinfectant wipes against bacteria cells (human lung epithelial carcinoma cells) which origi- and C. albicans . Briefly, four squares as test fields nated from the Institute of Medical Virology, Hannover were marked on a PVC with PUR surface coating material Medical School. Polyomavirus SV40 strain 777 was propa- (20 cm × 50 cm), figuring a row at a distance of 7 cm from gated in CV-1 cells (kidney cells of African green monkey, one another (Fig. 1). The marked test field 1 on this floor- Friedrich-Loeffler-Institute RIE 185). The Minimum Essen- ing was inoculated with the inoculum based on the test tial Medium was supplied by Lonza Verviers, Belgium and virus suspension and a solution of interfering substance the fetal calf serum (FCS) by Biochrom GmbH, Germany. (clean conditions, 0.3 g/L BSA). Here, 50 μl inoculum was pipetted on the first test field (field 1) and distributed with Wipes a glass spatula. Immediately after drying of the inoculum Four commercial disinfectant wipes were examined in on test field 1 at 20 °C – 25 °C the wipe was fixed under a the 4-field test. Wipe A is a disinfectant wipe based on unitary weight (granite block with a weight between 2.3– 0.06% per acetic acid with a bactericidal, virucidal and 2.5 kg). This unitary weight should simulate the average sporicidal claim. Wipe B is based on 0.6% quaternary pressure during the wiping process. For the examination ammonium compounds (QACs) with a claim against the granite block with the fixed wipe was rapidly moved bacteria, spores and viruses. The active ingredients of from test field 1 to test field 4 and back within no longer wipe C are 0.78% QACs with a claim against bacteria, than 2 s. At the end of the contact time (5 min chosen for enveloped viruses, norovirus and SV40. Wipe D is based all experiments) the test organisms were recovered from on 70% (v/v) 2-propanol with a use mainly in clean all four fields with a moistened and a dry nylon swab rooms and with a claim against bacteria only. (FloQSwab, art. no. 529CS0, Copan Diagnostics Inc., Man- In addition, the active solutions of all wipes were exam- tua, Italy) as described in the EN 16615:2015 . Swabs ined in combination with a reference wipe. The reference of each field were transferred to 5 ml Minimum Essential wipe was the Tork Standard wipe, art. no. 90491 supplied Medium (MEM), respectively and tubes were vortexed for by SCA Tork (D-68305 Mannheim) (17.5 cm × 28 cm, 60 s. Virus titres of the eluates were determined immedi- 55% pulp, 45% polyethylenterephthalat) as described in ately by end point dilution techniques as described in the the EN 16615:2015 . All experiments were performed EN 14476:2015  and calculated using the method of in two independent runs. Kärber  and Spearman . The virus titre is expressed as log TCID /ml with 95% confidence inter- 10 50 4-field test with viruses val. The virus reduction was calculated by comparing the The performance of the 4-field test is described in detail virus titres of each test field with those immediately after in the EN 16615:2015 as European Standard for drying and the chosen exposure time. Fig. 1 PVC material with polyurethane surface coating measuring 20 × 50 cm is prepared by recording four 5 × 5 cm squares. Test field 1 is contaminated with a defined amount of virus inoculum (virus suspension + interfering substance). This is followed by a wiping process (2-s motion) with a wipe, which was fixed under a unitary weight. After a defined exposure time of the test substance, the remaining test viruses are recovered from all four test fields with swabs. The aim is to show a reduction of the test viruses on test field 1 and the carryover to the previously virus-free test fields 2–4 (schematic drawing according to European Standard EN 16615:2015) Becker et al. Antimicrobial Resistance and Infection Control (2019) 8:121 Page 4 of 8 Controls in the 4-field test with viruses causing false-positive results, 1.25 ml of the resulting The following controls were included: eluate of the cytotoxicity control was contaminated with 12.5 μl test virus suspension and stored for 30 min. Fi- Initial virus control (VIC) nally, the virus titre was determined by end point dilu- For calculating the initial virus titre 0.05 ml inoculum tion method. For test validation the difference between was mixed with 5 ml MEM followed by determination of NC and VIC should be ≤0.5 log steps as described in the virus titre by end point dilution titration on permis- the EN 14476:2015 . sive cells. Interference control (IC) Drying control Here it must be excluded that the residual product in Drying controls were performed immediately after dry- the eluate will influence the infectivity of the cells and ing of the virus inoculum on a test field (DCt0) and after thus might inhibit the virus propagation causing false- the defined exposure time of 5 min (DCt5) with the positive results. 2.5 ml of the eluate of the cytotoxicity same recovery procedure as described above. The DCt0 control or phosphate-buffered saline (PBS) was mixed shows the loss of virus during the drying process. DCt5 with 2.5 ml of a double-concentrated cell suspension and was the reference for calculation of the reduction factor stored for 1 h at 37 °C. Afterwards the cells were re- (RF) on the test fields 1–4, respectively. suspended and the virus suspension was titrated with these cells. The PBS treated cell suspension served as a Water control control. For validation of the test the difference between To discriminate the virus-inactivation properties of the both assays should be lower than 1.0 log steps as de- wipes from a mechanical effect on test field 1 two water scribed in the EN 14476:2015 . controls (water of standardized hardness (WSH) and Aqua bidest.) with the Tork Standard wipe were included. Virus detection in the wipes after usage In addition, the virus titres were measured on test fields The wipes were examined for virus after the usage. Here, 2–4 for control of the virus transfer with water. the area used for wiping was cut out and this material was transferred in a 50 ml tube with 10 ml MEM with- Cytotoxicity control out fetal calf serum (FCS). After vortexing for 30 s and a Test field 1 was inoculated with an inoculum with MEM squeezing-out step with a sterile glass spatula 100 μlof instead of virus suspension. A run with the respective each fluid was analysed for virus by microtitration on wipe and the elution followed. Finally, the eluate of test the appropriate cell lines. field 1 was added to the corresponding cell cultures. This cytotoxicity control defines the lower detection Results limit of the test system for the corresponding wipe. Performance test control measures In the beginning, the virus titre of the inoculum (VIC) Neutralisation control (NC) was compared with the titres on the PVC flooring imme- To exclude that the resulting eluate after immediate di- diately after being visibly dry (DCt0) and after 5 min ex- lution is still expressing any virus-inactivating property posure time (DCt5). Results are shown in Fig. 2. SV40 was Fig. 2 Stability of three test viruses adenovirus (AdV) type 5, murine norovirus (MNV) and polyomavirus SV40 (SV40) under clean conditions immediately after drying (DCt0) and after 5 min exposure time (DCt5) in comparison to the inoculum without drying (VIC). The calculated reductions of virus titre after 5 min were 0.34 for SV40, 1.47 for MNV and 2.04 for AdV Becker et al. Antimicrobial Resistance and Infection Control (2019) 8:121 Page 5 of 8 the most stable virus (reduction 0.15 log steps after dry- test field 1 against all three test viruses. In contrast, wipe ing) followed by MNV (1.16) and AdV (2.07). The add- B and C were not able to inactivate sufficiently MNV itional exposure time of 5 min only produced minor and AdV on test field 1. However both QAC-based changes of the virus reduction for all three test viruses. wipes were active against SV40 (≥ four log steps on With the water control the transfer of the dried virus test field 1). The 2-propanol-based wipe (wipe D) was from test field 1 to the other test fields is visible. The not able in inactivate any of the three test viruses to the reference wipe Tork Premium Spezial Tuch was treated desired extent. with water of standardized hardness (WSH) and with The results with the pre-wetted wipes and the active Aqua bidest. in parallel. After wiping and the chosen ex- fluids tested with the Standard Tork wipe were nearly posure time, virus titres on test fields 1–4 of the water identical (Fig. 4). Only the results of wipe B and the cor- controls were detected. Additionally, virus titres of DCt0 responding active solution were different. The active so- and DCt5 were measured (Fig. 3). lution of wipe B (QAC-based formulation) was effective Examining MNV the calculated loss of virus titre after against MNV resulting in a four log reduction (RF = the drying step and water treatment causes RFs of 2.88 4.19), whereas the corresponding wipe failed (RF = 3.44) (WSH) and 3.00 (Aqua bidest.). This virus loss was the thus producing a nearly identical RF than WSH (RF = highest among the three test viruses followed by AdV. 2.88) and Aqua dest. (RF = 3.00). Here the RFs were 2.50 (WSH) and 2.63 (Aqua bidest.). A remarkable transfer to fields 2–4 was only seen When testing SV40 only a small reduction of virus titre when testing the 2-propanol-based wipe D with all three was observed after drying and water treatment (RFs = viruses (Table 1). Additionally, SV40 was transferred to 1.44 with WSH and 2.00 with Aqua bidest). In contrast, test fields 2–4 by the product C. In all other experiments the highest titres on the other test fields were observed no transfer of viruses from test field 1 to the other fields with SV40 demonstrating a great transfer of this virus to was measured (Table 1). test fields 2–4. With AdV and MNV only lower virus ti- tres were measured on test fields 2–4 (Fig. 3). In the EN Examination of the wipes for viral contamination 16615:2015 it is requested to demonstrate in the water Next, the wipes were examined for contamination control a transfer of bacteria and C. albicans to the test with viral material. Figure 5 shows that in the PAA- fields 2–4 which is also shown here with the test vi- based wipe A no residual virus could be detected ruses. Concerning removal from test field 1 and transfer after usage. In all other wipes and in the Standard to test fields 2–4 resulting data with WSH and Aqua wipe treated with Aqua bidest. MNV, AdV and SV40 bidest. were nearly identical. couldbedetected(Fig. 5). Virucidal efficacy of the wipes Discussion The four wipes exhibited different virucidal efficacies Virus transfer in the hospital can be interrupted by the against the chosen viruses (Fig. 4). The PAA-based wipe appropriate cleaning and disinfection of surfaces. Pre- (wipe A) was able to achieve a four log reduction on wetted wipes may play a role in this . Meanwhile, Fig. 3 The Tork Standard wipes were treated with water of standardized hardness (WSH) and Aqua bidest. (Aqua) for studying the mechanical removal from test field 1 and the transfer to test fields 2–4 with AdV, MNV and SV40. The titres of the inoculum, after drying (DCt0) and 5 min exposure time (DCt5) together with titres on test fields 1–4 and a summary of transfer to test fields 2–4 (accumulation factor (AF) fields 2–4) are shown. The lower detection limit defined by the cytotoxicity is indicated by a dashed line Becker et al. Antimicrobial Resistance and Infection Control (2019) 8:121 Page 6 of 8 Fig. 4 Virucidal properties (reduction factor on field 1) of four commercial wipes and the corresponding fluids in comparison with WSH and Aqua bidest. Against AdV, MNV and SV40 in the 4-field test. The left columns (light grey) show the efficacy of the fluids with the Tork Standard wipe and the right columns (dark grey) of the commercial disinfectant wipes by giving the reduction factor (RF). A four log reduction (inactivation 99.99%) is considered for efficacy detergent and disinfectant wipes with a proven efficacy Our choice of the test virus was mainly influenced by against bacteria and C. albicans are available based on existing suspension tests [16, 17]. In addition, AdV and data of the 4-field test described in the EN 16615:2015 MNV as a surrogate of human norovirus are also test vi-  or the ASTM E2967–15 . ruses in the prEN 16777:2016  and the German The virucidal claim of these pre-wetted wipes in Eur- Guideline of DVV . SV40 is a test virus in the Ger- ope is nowadays nearly completely based on quantitative man Guideline of DVV/RKI for testing disinfectants in suspension tests only like the EN 14476:2015  or the suspension and was introduced in the past as a surrogate German Guideline of DVV and RKI . In some cases of polyomaviruses . Therefore, tests with mechanical a test simulating practical conditions without mechanical action should include viruses that have been studied in action is performed in addition . However, a claim quantitative suspension tests and/or in carrier tests with- against viruses with a practical test like the 4-field test out mechanical action. with mechanical action including inactivation and re- First of all the viral stability of the three test viruses moval steps would provide more precise information for was examined after drying. SV40 was more stable during these pre-wetted wipes used in healthcare. this process than MNV and AdV. The greatest decline of Meanwhile, there are two standards designed to meas- virus titre was found with AdV during the drying stud- ure the claims of disinfectant pre-wetted wipes. We have ies. However, in summary, despite the drop in virus titre chosen the EN 16615:2015  in contrast to the ASTM with all chosen test viruses it is still possible to demon- E2967–15  with the Wiperator due to the possibility strate a four log reduction later in the tests with the to check the transfer of bioburden in one process and wipes necessary for claiming a sufficient efficacy. In con- the shorter wiping procedure. The EN 16615:2015 de- trast to bacteria testing four log reduction is necessary scribes a manual horizontal movement (2 s) in contrast in virus testing as described in the DIN EN 14476:2015 to the orbital mechanical rotation (10 s) with the  due to the fact that high titers in many cases are dif- Wiperator. The possible transfer in the EN 16615:2015 ficult to reach and that high cytotoxicity of the disinfec- can be checked when examining the virus load on test tants will increase the lower detection limit. fields 2–4. In contrast, only when introducing an add- Tests with WSH and Aqua bidest showed that the loss itional step the Wiperator provides information on the of virus titre on test field 1 was the greatest with MNV bacterial transfer from the wipe to three consecutive followed by AdV and SV40. With SV40 there was only a stainless disks together with the effect of the mechanical small loss by drying but a great virus transfer to the action (10 s, 150 g pressure) . other test fields whereas with MNV and AdV only a Table 1 Transfer of the three test viruses AdV, MNV and SV40 to fields 2, 3 and 4 when using the four different wipes. The virus titres are given as TCID /ml on test fields 2–4 (CT = cytotoxicity, no virus = no virus detected) AdV MNV SV40 field 2 field 3 field 4 field 2 field 3 field 4 field 2 field 3 field 4 Aqua 1.33 ± 0.13 1.33 ± 0.13 1.23 ± 0.04 1.39 ± 0.07 1.33 ± 0.07 1.20 ± 0.00 4.42 ± 0.10 4.11 ± 0.10 3.70 ± 0.19 Wipe A no virus no virus no virus no virus no virus no virus no virus no virus no virus Wipe B no virus no virus no virus no virus no virus no virus no virus no virus no virus Wipe C no virus no virus no virus no virus no virus no virus 3.45 ± 0.25 3.32 ± 0.13 3.76 ± 0.07 Wipe D 1.51 ± 0.06 1.51 ± 0.19 1.57 ± 0.38 1.76 ± 0.44 1.51 ± 0.06 1.39 ± 0.06 4.70 ± 0.13 4.95 ± 0.13 4.57 ± 0.38 Becker et al. Antimicrobial Resistance and Infection Control (2019) 8:121 Page 7 of 8 Fig. 5 Determination of residual virus in the four wipes (a-d). Virus titres are given as log TCID in 10 ml. No residual virus was detectable in 10 50 the PAA-based wipe (wipe A). The lower detection limit is here defined by the cytotoxicity small transfer resulted. This means when using wipes comparison to the detergent wipes examined with vege- the transfer to consecutive surfaces might be influenced tative bacteria in the other study . by the kind of virus contamination. The great efficacy of the PAA-based wipe A in the 4- Testing the different wipes the greatest reduction of field test was finally confirmed by the examination of virus titre was measured with the PAA-based wipe A the wipes after usage. None the three test viruses could resulting in greater than four log steps against all three be detected in this wipe in contrast to the others. test viruses. In general, a four log reduction of titre in virus tests is necessary for claiming efficacy as in other Conclusions virucidal test methods. Despite the fact, that wipe B, un- In summary, we showed that the principle of the existing like the active solution, just missed a four log reduc- EN 16615:2015 can be transferred to the examination tion, nearly identical results were found when testing the with viruses. Our data demonstrate that a successful pre-wetted wipes and the corresponding fluids with the virus inactivation and a prevention of virus transfer can Tork Standard wipe. This means that the material of the be reached. Consequently, a 4-field test evaluating viru- wipes used in this study seemed to have no great impact cidal activity of disinfectant wipes is possible and will on efficacy. Identical results were found by Wesgate and allow more precise information for virucidal claims of co-workers when examining different products with wipes. microfiber, cotton or non-woven materials . These Besides the possibility to introduce wipes with a viru- examinations were performed with Pseudomonas aerugi- cidal claim the appropriate handling “one site-, one dir- nosa and S. aureus and different disinfectant solutions ection, one use” is still of great importance for hospital by the ASTM method and the EN 16615:2015. hygiene. Therefore, this correct management of the The QAC-based wipes B and C were also able to reach wipes together with an appropriate claim will help to efficacy four log reduction against SV40 but failed to control viral bioburden on healthcare surfaces. In inactivate MNV and AdV. The 2-propanol based prod- addition, future studies have to address the area of activ- uct (wipe D) was inactive against all three chosen test vi- ity of the chosen disinfectant wipe in comparison to de- ruses. These marked differences in efficacy of wipes tergent wipes. Finally, the detection of all test viruses in should also be observed when testing with bacteria . three of four wipes tested makes a careful disposal of However in a study examining detergent wipes with S. great importance. aureus, A. baumannii and C. difficile all seven detergent wipes were not able to produce a sufficient reduction and all wipes transferred significant amounts of bacteria Abbreviations AdV: Adenovirus; ASTM: American Society for Testing and Materials; and spores to consecutive surfaces . In our study, DVV: Deutsche Vereinigung zur Bekämpfung der Viruskrankheiten e.V.; the disinfectant wipes under examination were mainly EN: European Norm; HITES: High touch environmental surfaces; chosen with respect to their ability to already inactivate MEM: Minimum Essential Medium; MNV: Murine norovirus; PAA: Per acetic acid; PBS: Buffered-saline solution; PCR: Polymerase chain reaction; microorganisms including different viruses in suspension QAC: Quaternary ammonium compound; RF: Reduction factor; RKI: Robert assays. By doing so, a virucidal activity in the 4-field test Koch-Institute; RV-A: Group A rotavirus; SV40: Polyomavirus SV40; WSH: Water was at least expected for disinfectant wipes in of standardized hardness Becker et al. Antimicrobial Resistance and Infection Control (2019) 8:121 Page 8 of 8 Acknowledgements 10. Boyce JM. Modern technologies for improving cleaning and disinfection of We acknowledge support by the DFG Open Access Publication Funds of the environmental surfaces in hospitals. Antimicrob Resist Infect Control. 2016;5: Ruhr University Bochum. 10. 11. Siani H, Wesgate R, Maillard JY. Impact of antimicrobial wipes compared with hypochlorite solution on environmental surface contamination in a Authors’ contributions health care setting: a double-crossover study. Am J Infect Control. 2018;46: All authors contributed to the manuscript according to the ICMJE 1180–7. [International Committee of Medical Journal Editors] recommendations: LH 12. DIN EN 16615:2015. Chemical disinfectants and antiseptics. Quantitative test and BB were responsible for data acquisition. All authors were involved in method for the evaluation of bactericidal and yeasticidal activity on non- analysis and interpretations of the data. JS and ES prepared the manuscript. porous surfaces with mechanical action employing wipes in the medical All authors critically revised the manuscript and are accountable for accuracy area [4-field test]. Test method and requirements [phase 2, step 2]. Brussels and correctness. All authors have read and agreed to the final draft before 2015, CEN-Comité Europeen de Normalisation. submission. 13. ASTM E2967-15. Standard test method for assessing the ability of pre- wetted towelettes to remove and transfer bacterial contamination on hard, Funding non-porous environmental surfaces using the Wiperator. West This research was supported by the Deutsche Gesellschaft für Conshohocken 2015, PA ASTM International 2015. Krankenhaushygiene. 14. Sattar SA, Bradley C, Kibbee R, Wesgate R, Wilkinson MA, Sharpe T, Maillard JY. 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