この作業は、米国防災庁（AID-OFDA-A-15-00026）によって支援された。 Marlene WolfeはNational Science Foundation（助成金0966093）の支援を受けました。

<ol>
	<li>Kampf, G., Kramer, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiologic+Background+of+Hand+Hygiene+and+Evaluation+of+the+Most+Important+Agents+for+Scrubs+and+Rubs.">Epidemiologic Background of Hand Hygiene and Evaluation of the Most Important Agents for Scrubs and Rubs.</a> Clin Microbiol Rev. 17 (4), 863-893 (2004).</li><li>Miller, T., Patrick, D., Ormrod, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hand+decontamination:+influence+of+common+variables+on+hand-washing+efficiency.">Hand decontamination: influence of common variables on hand-washing efficiency.</a> Healthc Infect. 16 (1), 18 (2013).</li><li>Jensen, D. A., Danyluk, M. D., Harris, L. J., Schaffner, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantifying+the+effect+of+hand+wash+duration,+soap+use,+ground+beef+debris,+and+drying+methods+on+the+removal+of+Enterobacter+aerogenes+on+hands.">Quantifying the effect of hand wash duration, soap use, ground beef debris, and drying methods on the removal of Enterobacter aerogenes on hands.</a> J Food Prot. 78 (4), 685-690 (2015).</li><li>Girou, E., Loyeau, S., Legrand, P., Oppein, F., Brun-Buisson, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+of+handrubbing+with+alcohol+based+solution+versus+standard+handwashing+with+antiseptic+soap:+randomised+clinical+trial.">Efficacy of handrubbing with alcohol based solution versus standard handwashing with antiseptic soap: randomised clinical trial.</a> BMJ. 325 (7360), 362 (2002).</li><li>Kac, G., Podglajen, I., Gueneret, M., Vaupr&#233;, S., Bissery, A., Meyer, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microbiological+evaluation+of+two+hand+hygiene+procedures+achieved+by+healthcare+workers+during+routine+patient+care:+a+randomized+study.">Microbiological evaluation of two hand hygiene procedures achieved by healthcare workers during routine patient care: a randomized study.</a> J Hosp Infect. 60 (1), 32-39 (2005).</li><li>Lages, S. L. S., Ramakrishnan, M. A., Goyal, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In-vivo+efficacy+of+hand+sanitisers+against+feline+calicivirus:+a+surrogate+for+norovirus.">In-vivo efficacy of hand sanitisers against feline calicivirus: a surrogate for norovirus.</a> J Hosp Infect. 68 (2), 159-163 (2008).</li><li>Holton, R. H., Huber, M. A., Terezhalmy, G. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antimicrobial+efficacy+of+soap+and+water+hand+washing+versus+an+alcohol-based+hand+cleanser.">Antimicrobial efficacy of soap and water hand washing versus an alcohol-based hand cleanser.</a> Tex Dent J. 126 (12), 1175-1180 (2009).</li><li>Salmon, S., Truong, A. T., Nguyen, V. H., Pittet, D., McLaws, M. -. 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H., Duizer, E., Beumer, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reducing+viral+contamination+from+finger+pads:+handwashing+is+more+effective+than+alcohol-based+hand+disinfectants.">Reducing viral contamination from finger pads: handwashing is more effective than alcohol-based hand disinfectants.</a> J Hosp Infect. 90 (3), 226-234 (2015).</li><li>Steinmann, J., Paulmann, D., Becker, B., Bischoff, B., Steinmann, E., Steinmann, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+virucidal+activity+of+alcohol-based+hand+sanitizers+versus+antimicrobial+hand+soaps+in+vitro+and+in+vivo.">Comparison of virucidal activity of alcohol-based hand sanitizers versus antimicrobial hand soaps in vitro and in vivo.</a> J Hosp Infect. 82 (4), 277-280 (2012).</li><li>de Aceituno, A. F., Bartz, F. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ability+of+Hand+Hygiene+Interventions+Using+Alcohol-Based+Hand+Sanitizers+and+Soap+To+Reduce+Microbial+Load+on+Farmworker+Hands+Soiled+during+Harvest.">Ability of Hand Hygiene Interventions Using Alcohol-Based Hand Sanitizers and Soap To Reduce Microbial Load on Farmworker Hands Soiled during Harvest.</a> J Food Protect. 78 (11), 2024-2032 (2015).</li><li>Boyce, J. M., Pittet, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=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.">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.</a> Infect Control Hosp Epidemiol. 23 (12 Suppl), S3-S40 (2002).</li><li>. UNDP Medical Waste Experts Assessment and Recommendations Regarding Management of Ebola-Contaminated Waste Available from: <a target="_blank" href="https://noharm-global.org/sites/default/files/documents-files/3127/Report%20to%20WHO%20WASH%20and%20Geneva%20on%20Ebola%20final.pdf">https://noharm-global.org/sites/default/files/documents-files/3127/Report%20to%20WHO%20WASH%20and%20Geneva%20on%20Ebola%20final.pdf</a> (2015)</li><li>Hopman, J., Kubilay, Z., Allen, T., Edrees, H., Pittet, D., Allegranzi, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+of+chlorine+solutions+used+for+hand+hygiene+and+gloves+disinfection+in+Ebola+settings:+a+systematic+review.">Efficacy of chlorine solutions used for hand hygiene and gloves disinfection in Ebola settings: a systematic review.</a> Antimicrob Resist Infect Control. 4 (1), 1 (2015).</li><li>Lowbury, E. J. L., Lilly, H. A., Bull, J. 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R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+eluents+for+the+recovery+of+an+enveloped+virus+from+hands+by+whole-hand+sampling.">Evaluation of eluents for the recovery of an enveloped virus from hands by whole-hand sampling.</a> J Appl Microbiol. 118 (5), 1210-1216 (2015).</li><li>Sinclair, R. G., Rose, J. B., Hashsham, S. A., Gerba, C. P., Haas, C. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Criteria+for+Selection+of+Surrogates+Used+To+Study+the+Fate+and+Control+of+Pathogens+in+the+Environment.">Criteria for Selection of Surrogates Used To Study the Fate and Control of Pathogens in the Environment.</a> Appl Environ Microbiol. 78 (6), 1969-1977 (2012).</li><li>Held, E., Skoet, R., Johansen, J. 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著者らは、競合する金銭的利益がないと宣言している。

The method described here provides an approach for testing handwashing efficacy in a controlled laboratory setting. This method highlights the use of human volunteers and surrogate, non-infectious organisms. Using the method, it was possible to demonstrate differences in: 1) the efficacy of handwashing methods and 2) organism persistence in rinse water. The purpose of presenting this protocol is to provide a general framework that can be adapted to test a wide range of surrogate organisms and handwashing methods relevant to infectious disease.
During the use of the method, two key data quality recommendations were noted as important. First, the inoculate must be applied both as similarly as possible across the rounds of testing and in a manner, that minimizes loss. This is to ensure that sufficient inoculate is applied to the hands to allow for statistically significant results. Second, be sure to complete the "cleansing wash" step, in which the protocol is performed without handwashing prior to testing, as previous work has shown that there are likely to be significant differences between a first wash and subsequent washes, but not between subsequent washes performed after a cleansing round29. Additionally, this step clears residual hand contamination, which would interfere with results. 
The main limitation of this protocol is that it can be uncomfortable for volunteers. During each round of testing, which lasted about 2 h, volunteers' hands became cold. Some volunteers reported mild pain from their hands being constantly wet. Additionally, after a few rounds of testing, volunteers' hands became supersaturated, no longer fully drying between rounds. Although the randomization of the order of handwashing methods for each volunteer accounted for supersaturation, it is possible that the supersaturation could act as a confounding or modifying factor in this type of testing. To address this limitation, it is recommended that volunteers are appraised of this risk during consent disclosures and are reminded of their right to drop out of the study at any time. Volunteers should not undergo testing for more than 2 h per day to allow time for the hands to return to a baseline state and to minimize discomfort. A second limitation is the need to use a surrogate organism or non-infectious variant of a pathogenic organism to protect the health of volunteers. This might cause concern about the generalizability of results. However, for some pathogenic organisms (such as the Ebola virus), this limitation cannot be ethically overcome. Care must be taken during surrogate organism selection. Lastly, this is a laboratory study on efficacy. Results may only translate to effective disease prevention in real-word contexts where handwashing methods are made accessible to those in need and are used properly and consistently.
This protocol draws on previous work on handwashing efficacy but attempts to streamline methods and emphasizes the use of human hands (rather than surrogate surfaces) for testing. Additionally, rinse water is a transmission risk that had previously not been assessed. Existing studies on handwashing efficacy vary in methodology, leading to non-comparable data. We hope that standardized protocols for conducting handwashing method comparisons will encourage comparable and replicable results. Previous work has demonstrated that in vitro testing on surrogate surfaces such as pig skin, where, for example, the actual Ebola virus could be used, produced results that do not match those found after testing on human hands38. Therefore, a method using human hands and surrogate or non-infectious organisms is currently the best available approach to estimate handwashing efficacy and rinse water persistence for infectious microorganisms.
Handwashing is critical to prevent disease transmission. However, there is a lack of evidence on the comparative efficacy of handwashing methods that are commonly recommended. This protocol can be used to generate evidence about handwashing efficacy and rinse water persistence. This is especially important for infectious diseases with the potential to cause large outbreaks, such as the Ebola virus. We hope that other researchers will find this protocol useful to generate much-needed additional evidence on handwashing method efficacy and rinse water persistence that will assist in developing recommendations to reduce the transmission of infectious diseases.

手洗いは、下痢や呼吸器疾患を含む、特に糞便 - 口腔または空中経路によって伝播される病気の拡大を防ぐために広く推奨されている1 。驚くべきことに、石けんと水（HWWS）による手洗いとアルコールベースの手指消毒剤（ABHS）による手洗い方法の有効性に関する比較可能な証拠はほとんどなく、生物を手から取り除く。初期の研究では、手洗い法とは対照的に手洗いの機械的作用が、ほとんどの生物の除去の原因となる可能性があることが分かっている2,3。さらに、手洗い法が最も効果的であるという比較の証拠はほとんどない。非公式の文献レビューでは、石鹸と手の消毒剤の有効性を生物の除去に比較した14の研究が確認された。これらの研究のうち、5人はABHSがより有効であることを見いだした4 、 </HWWSが9,10,11,12,13,14,15,12,13,14,15,16,17の間で有意差がないことが判明した。これらの知見は矛盾しており、手洗い後のすすぎ水中の生物の存続による疾病の継続的なリスクには対処していない。全体として、感染症を引き起こす病原体を除去するための手洗い方法の比較有効性に関する証拠は限られている。 この限られた証拠は、どの方法がアウトブレイクの状況に最も適切かについて不確実性をもたらしている。例えば2013年から2016年にかけての西アフリカでのエボラウイルス病（EVD）発生時に、いくつかの大規模な国際的対応者が、HWWS、ABHS、または0.05％塩素溶液に関する矛盾した推奨を提供した。世界保健機構（WHO）はHWWSまたはABHS（手が目に見えて汚れていない場合）を推奨しているが、手洗いには0.05％塩素溶液を使用することをMSCは推奨している。 WHOは、他の選択肢がない限り、塩素を使用すべきでないと述べている。なぜなら、それは皮膚18,19,20,21,22によって発せられる塩素要求のため他の方法よりも効果が低いからである。さらに、塩素溶液は、一般に、高濃度の次亜塩素酸塩（HTH）、局所的に生成され安定化された次亜塩素酸ナトリウム（NaOCl）、および苛性ソーダ（ジクロロイソシアヌレート（NaDCC））。西アフリカでのEVD流行に対応してWHOによって委託された体系的なレビューでは、最近、手洗いと塩素による比較の有効性を調査したわずか4件の研究しか見つかりませんでした23 。これらの研究でも矛盾する結果が得られましたが、これらの研究では、エボラウイルス10,24,25,26,27に類似した手洗いまたは調査された微生物について、0.05％の推奨塩素濃度を使用していませんでした。したがって、勧告はエビデンスベースであるとは認められず、どの勧告が最も効果的であるかは不明であった。 手洗いの介入は流行病の伝染を予防するための重要なツールであるため、感染病原菌の感染を予防するための手洗いアプローチを比較するためには、さらなる研究が必要です。これらのh洗濯推奨は証拠に基づいていなければならない。したがって、代理人または非感染性病原体を用いて実施される手洗い効力およびすすぎ水の持続性を試験する方法が開発された2,28,29。エボラウイルスの代理としてPhi6を使用し、共通指標生物として大腸菌（Escherichia coli）を使用した試料の結果をここに示す。このプロトコールでは、手洗いの有効性およびすすぎ水の持続性試験が提示される。

<table><tbody><tr><td>Soap bar</td><td>Dove</td><td>White Beauty Bar soap</td><td></td></tr><tr><td>Alcohol-based hand sanitizer</td><td>Purell</td><td>Advanced Instant Hand Sanitizer with 70% Ethyl Alcohol</td><td></td></tr><tr><td>HTH Powder</td><td>Acros Organics</td><td>300340010</td><td></td></tr><tr><td>NaDCC Powder</td><td>Medentech</td><td>Klorsept granules</td><td></td></tr><tr><td>NaOCl Solution</td><td>Acros Organics</td><td>419550010</td><td></td></tr><tr><td>Electrochlorinator</td><td>AquaChlor</td><td></td><td></td></tr><tr><td>Iodometric titrator</td><td>Hach</td><td>1690001</td><td></td></tr><tr><td>Bovine serum albumin</td><td>MP Biomedicals</td><td>NC0117242</td><td></td></tr><tr><td>Tryptone</td><td>Fisher</td><td>BP1421-100</td><td></td></tr><tr><td>Bovine Mucin</td><td>EMD Milipore</td><td>49-964-3500MG</td><td></td></tr><tr><td>0.22 &amp;micro;m Filter</td><td>EMD Milipore</td><td>GVWP04700</td><td></td></tr><tr><td>NaCl</td><td>Fisher</td><td>BP358-1</td><td></td></tr><tr><td>Skin pH probe</td><td>Hanna Instruments</td><td>H199181</td><td></td></tr><tr><td>Large Whirlpak Sample Bag</td><td>Nasco</td><td>B01447WA</td><td></td></tr><tr><td>Small Whirlpak Sample Bag</td><td>Nasco</td><td>B01323WA</td><td></td></tr><tr><td>Funnel bottle</td><td>Thermo Scientific</td><td>3120850001</td><td>You may drill an appropriately sized hole in the lid of a bottle to form a funnel that will dispense water at the appropriate flow rate</td></tr><tr><td>Ethanol</td><td>ThermoScientific</td><td>615090010</td><td>Mix with water to produce 70% ethanol</td></tr><tr><td>Spray bottle</td><td>Qorpak</td><td>PLC06934</td><td></td></tr><tr><td>&lt;em&gt;E. coli&lt;/em&gt;</td><td>ATCC</td><td>25922</td><td></td></tr><tr><td>LB Broth</td><td>Fisher BioReagents</td><td>BP1426-2</td><td></td></tr><tr><td>LB Agar</td><td>Fisher BioReagents</td><td>BP1425-500</td><td></td></tr><tr><td>Sterile loop</td><td>Globe Scientific</td><td>22-170-204</td><td></td></tr><tr><td>Phi6</td><td>HER</td><td>102</td><td></td></tr><tr><td>Nutrient broth</td><td>BD Difco</td><td>BD 247110</td><td></td></tr><tr><td>GeneQuant 100 Spectrophotometer</td><td>General Electric</td><td>28-9182-04</td><td></td></tr><tr><td>Sodium thiosulfate</td><td>Fisher Chemical</td><td>S445-3</td><td></td></tr><tr><td>Membrane filter (47 mm, 0.45 &amp;micro;m)</td><td>EMD Millipore</td><td>HAWP04700</td><td></td></tr><tr><td>m-ColiBlue24 broth media</td><td>EMD Millipore</td><td>M00PMCB24</td><td></td></tr><tr><td>Petri dish with pad (47 mm)</td><td>Fisherbrand</td><td>09-720-500</td><td></td></tr><tr><td>Vacuum Manifold</td><td>Thermo Scientific/Nalgene</td><td>09-752-5</td><td></td></tr><tr><td>Filter funnels</td><td>Thermo Scientific/Nalgene</td><td>09-747</td><td></td></tr><tr><td>Pseudomonas syringae</td><td>HER</td><td>1102</td><td></td></tr><tr><td>Phosphate Buffered Saline</td><td>Thermo Scientific</td><td>10010031</td><td>Solution may also be mixed from source compounds according to any basic recipe</td></tr></tbody></table>

a method to test the efficacy of handwashing for the removal of emerging infectious pathogens

感染症の伝染を防ぐために手洗いが広く推奨されています。しかしながら、一般的な手洗い方法の有効性については、匹敵する証拠はほとんど存在しない。さらに、手洗い法を比較して、感染性病原体を除去するのに最も効果的なものを決定する証拠はほとんどない。感染症発生時に使用される可能性のある手洗いのさまざまなアプローチの証拠を提供するための研究が必要です。ここでは、手から微生物を除去する際の手洗い方法の有効性およびすすぎ水中でのその持続性を評価するための実験方法が記載されている。ボランティアの手を最初に試験生物にスパイクし、次に各手洗い方法で洗浄する。一般に、代理微生物は、ヒト被験体を疾患から保護するために使用される。洗浄後にボランティアの手に残っている生物の数は、変更された「グローブジュース」法を用いて試験される。手はエルツ微生物を懸濁させ、膜濾過（バクテリア）またはプラークアッセイ（ウイルス/バクテリオファージ）による分析に利用できるようにスクラビングされる。手洗いから生成されたすすぎ水は、分析のために直接収集される。手洗い効果は、手洗い後に採取した試料と手洗いをしていない試料との対数減少値を比較することによって定量化する。すすぎ水の持続性は、様々な手洗い方法からのすすぎ水サンプルを、単に水で手洗いした後に採取したサンプルと比較することによって定量化される。この方法は、ヒトボランティアの安全性を維持するために代理生物を使用する必要性によって制限されるが、 インビトロ研究では複製が困難で手洗いの有効性に関する研究ギャップやリンスにおける感染性生物の持続性を補う水。

ここでは、プロトコル（ 図1 ）は18名のボランティアで完了し、各ボランティアは大腸菌とPhi6の両方を用いて試験した。土壌負荷の有無にかかわらず大腸菌による手洗い結果と土壌負荷によるPhi6との間に有意な差が見られた（ 図2および図3 ）。土壌負荷のない大腸菌では、HTH、NaDCC、および安定化NaOClを用いた手洗いのすべてが、水のみによる手洗い（F（6,102）= 2.72、p = 0.034）より有意に大きな対数減少をもたらした。土壌負荷では、HTHは水のみ、HWWSおよびABHS（F（6,102）= 3.94、p &lt;0.001）よりも大腸菌の対数減少が有意に大きかった。土壌負荷のないPhi6の方法間に有意差はなかった（F（6,66）= 2.04、p = 0.073）。しかし、土壌負荷のあるPhi6（F（6,102）= 7.01、p &lt;0.001）では、水だけでgreABHSまたは安定化されたNaOClよりも対数減少、およびHWWSは、ABHS、安定化されたNaOCl、および生成されたNaOClより大きな対数減少である。 HTHはまた、ABHSおよび安定化されたNaOClよりも大きな対数減少を有し、NaDCCは、安定化されたNaOClおよびABHSよりも大きな対数減少をもたらした。 HTHは条件全体で一貫して最も一貫して実行されましたが、多くの場合、0.5ログ未満から1.5ログ減少までの信頼区間が多いため、重要な結果の過剰解釈に対する注意が必要でした。 すすぎ水では、塩素はHWWSよりもすすぎ水に耐える大腸菌の対数減少が有意に大きかった（土壌負荷なし、F（4,68）= 331.7、p &lt;0.001;土壌負荷F（4,68） ）= 162.44、p &lt;0.001）（ 図4 ）。この同じパターンは、土壌負荷なしでPhi6で見られた（（F（4,43）= 8.95、P &lt;0.001）。すべての塩素溶液はすすぎ水中のPhi6の減少はHWWSよりも有意に大きい。 Phi6および土壌負荷（（F（4,67）= 3.35、p = 0.071）によるすすぎ水の持続性に有意差はなかった（ 図5 ）。 <img alt="図1" class="xfigimg" src="/files/ftp_upload/55604/55604fig1.jpg" /> 図1：実験の概要 1回のpH試験、2）手の接種、3）手洗い、4）手洗い、および5）試験した8つの条件のそれぞれについて手を汚染除去することが含まれる。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55604/55604fig1large.jpg" target="_blank">この図の拡大版を見るには、ここをクリックしてください。</a> <img alt="図2" class="xfigimg" src="/files/ftp_upload/55604/55604fig2.jpg" /> 図2： 大腸菌の手洗いのres<html>ウルトラ。手洗いをしない場合と比較して、手洗い法で試験した結果、土壌負荷なしで1.94-3.01、土壌負荷で2.18-3.34の大腸菌における平均対数減少がもたらされた 。水で手洗いすることは、両方の条件（1.94および2.18対数）において大腸菌の減少が最も少ないことを実証した 。 NaDCCによる手洗いは、土壌負荷なしで最大の減少（3.01）をもたらし、HTHは土壌負荷（3.34）で最大の減少をもたらした。チャートでは、線は微生物の減少率を表し、誤差バーは対数減少の標準誤差を表す。 Ctrl B、コントロールB; HWWS、石けんで手洗い; ABHS、アルコールベースの手指消毒剤。 HTH、高濃度の次亜塩素酸塩; NaDCC、ジクロロイソシアヌレートナトリウム; st NaOCl、安定化次亜塩素酸ナトリウム;次亜塩素酸ナトリウムを生成した。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55604/55604fig2large.jpg" target="_blank">この図の拡大版を見るには、ここをクリックしてください。</a> ve_content &quot;fo：keep-together.within-page =&quot; 1 &quot;&gt; <img alt="図3" class="xfigimg" src="/files/ftp_upload/55604/55604fig3.jpg" /> 図3：Phi6手洗いの結果。手洗いをしない場合と比較して、手洗い法では、Phi6の平均対数減少は土壌負荷なしで2.44-3.06、土壌負荷では2.71-3.69であった。石鹸を用いた手洗いは、土壌負荷なしでPhi6の減少を最小限に抑え（2.44）、安定化NaOClによる手洗いは土壌負荷（2.71）で最小の減少をもたらした。生成されたNaOClによる手洗いは土壌負荷なしで最大の減少（3.06）をもたらし、石鹸による手洗いは土壌負荷（3.69）で最大の減少をもたらした。チャートでは、線は微生物の減少率を表し、誤差バーは対数減少の標準誤差を表す。 Ctrl B、コントロールB; HWWS、石けんで手洗い; ABHS、アルコールベースの手指消毒剤。 HTH、高濃度の次亜塩素酸塩; NaDCC、ジクロロイソシアヌレートナトリウム;st NaOCl、安定化次亜塩素酸ナトリウム;次亜塩素酸ナトリウムを生成した。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55604/55604fig3large.jpg" target="_blank">この図の拡大版を見るには、ここをクリックしてください。</a> <img alt="図4" class="xfigimg" src="/files/ftp_upload/55604/55604fig4.jpg" /> 図4： 大腸菌の手洗い結果。水のみによる手洗いと比較して、すすぎ水に残っている大腸菌の平均対数減少は、土壌負荷なしでは0.28~4.77であり、土壌負荷では0.21~4.49であった。土壌負荷があってもなくても、石けんで手洗いで最小の減少が見られた（0.28および0.21）。最大の減少は、土壌負荷なしで安定して生成されたNaOCl（4.77）およびHTHおよび土壌負荷を伴うNaOClを伴って観察された。グラフでは、線は生物の減少率を表し、誤差棒はログ削減の標準誤差。 HWWS、石けんで手洗い; ABHS、アルコールベースの手指消毒剤。 HTH、高濃度の次亜塩素酸塩; NaDCC、ジクロロイソシアヌレートナトリウム; st NaOCl、安定化次亜塩素酸ナトリウム;次亜塩素酸ナトリウムを生成した。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55604/55604fig4large.jpg" target="_blank">この図の拡大版を見るには、ここをクリックしてください。</a> <img alt="図5" class="xfigimg" src="/files/ftp_upload/55604/55604fig5.jpg" /> 図5：Phi6ハンドリンス結果。すすぎ水に残っているPhi6の平均対数減少は、水のみによる手洗いと比較して、土壌負荷なしで1.26-2.02であり、土壌負荷では1.30-2.20であった。土壌負荷では、せっけん（1.26）による手洗いで最小の減少が見られた。土壌負荷がなければ、HTHは最小の減少（2.02）をもたらした。 NaDCCによる土壌負荷の有無にかかわらず、最大の減少が観察された（2.02および2.20）。チャートでは、線は微生物の減少率を表し、誤差バーは対数減少の標準誤差を表す。 HWWS、石けんで手洗い; ABHS、アルコールベースの手指消毒剤。 HTH、高濃度の次亜塩素酸塩; NaDCC、ジクロロイソシアヌレートナトリウム; st NaOCl、安定化次亜塩素酸ナトリウム;次亜塩素酸ナトリウムを生成した。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55604/55604fig5large.jpg" target="_blank">この図の拡大版を見るには、ここをクリックしてください。</a>

Watch this Scientific Journal Video about A Method to Test the Efficacy of Handwashing for the Removal of Emerging Infectious Pathogens at JoVE.com

A Method to Test the Efficacy of Handwashing for the Removal of Emerging Infectious Pathogens

感染症の伝染を防ぐために手洗いが広く推奨されています。しかし、どのような手洗い方法が感染症病原体を除去するのに最も効果的であるかに関する証拠はほとんどない。我々は、微生物を除去する手洗い方法の有効性を評価する方法を開発した。

発生する感染性病原体の除去のための手洗いの有効性を試験する方法

Handwashing is widely recommended to prevent infectious disease transmission. However, little comparable evidence exists on the efficacy of handwashing ...

a-method-to-test-efficacy-handwashing-for-removal-emerging-infectious

Nanyang Technological University

Research

JoVE Journal

Immunology and Infection

18.9K ビュー.  Tufts University. 感染症の伝染を防ぐために手洗いが広く推奨されています。しかし、どのような手洗い方法が感染症病原体を除去するのに最も効果的であるかに関する証拠はほとんどない。我々は、微生物を除去する手洗い方法の有効性を評価する方法を開発した。 

ビデオ: A Method to Test the Efficacy of Handwashing for the Removal of Emerging Infectious Pathogens

EPA Method 1615. Measurement of Enterovirus and Norovirus Occurrence in Water by Culture and RT-qPCR. I. Collection of Virus Samples

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. The standardized sampling component of the method concentrates viruses that may be present in water by passage of a minimum specified volume of water through an electropositive cartridge filter. The minimum specified volumes for surface and finished/ground water are 300 L and 1,500 L, respectively. A major method limitation is the tendency for the filters to clog before meeting the sample volume requirement. Studies using two different, but equivalent, cartridge filter options showed that filter clogging was a problem with 10% of the samples with one of the filter types compared to 6% with the other filter type. Clogging tends to increase with turbidity, but cannot be predicted based on turbidity measurements only. From a cost standpoint one of the filter options is preferable over the other, but the water quality and experience with the water system to be sampled should be taken into consideration in making filter selections.

EPA Method 1615 uses an electropositive filter to concentrate enteroviruses and noroviruses in environmental and drinking waters. This manuscript describes the procedure for collecting samples for Method 1615 analyses.

エンテロウイルスと文化およびRT-QPCRによる水中のノロウイルス発生のEPAメソッド1615測定。ウイルスサンプルのI.コレクション

EPA Method 1615 was developed with a goal of providing a standard method for measuring enteroviruses and noroviruses in environmental and drinking waters. ...

環境学

Determining Viral Disinfection Efficacy of Hot Water Laundering

This protocol provides an example of a laboratory process for conducting laundering studies that generate data on viral disinfection. While the protocol was developed for research during the coronavirus disease 2019 (COVID-19) pandemic, it is intended to be a framework, adaptable to other virus disinfection studies; it demonstrates the steps for preparing the test virus, inoculating the test material, assessing visual and integrity changes to the washed items due to the laundering process, and quantifying the reduction in viral load. Additionally, the protocol outlines the necessary quality control samples for ensuring the experiments are not biased by contamination and measurements/observations that should be recorded to track the material integrity of the personal protective equipment (PPE) items after multiple laundering cycles. The representative results presented with the protocol use the Phi6 bacteriophage inoculated onto cotton scrub, denim, and cotton face-covering materials and indicate that the hot water laundering and drying process achieved over a 3-log (99.9%) reduction in viral load for all samples (a 3-log reduction is the disinfectant performance metric in U.S. Environmental Protection Agency's Product Performance Test Guideline 810.2200). The reduction in viral load was uniform across different locations on the PPE items. The results of this viral disinfection efficacy testing protocol should help the scientific community explore the effectiveness of home laundering for other types of test viruses and laundering procedures.

In response to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, a laboratory protocol was developed to test the viral disinfection efficacy of hot water laundering of cloth face coverings, cotton scrubs, and denim pants. The Phi6 virus (bacteriophage) was used as the organism to test disinfection efficacy.

温水洗浄のウイルス消毒効果の決定

This protocol provides an example of a laboratory process for conducting laundering studies that generate data on viral disinfection. While the protocol ...

工学

Swab Sampling Method for the Detection of Human Norovirus on Surfaces

Human noroviruses are a leading cause of epidemic and sporadic gastroenteritis worldwide. Because most infections are either spread directly via the person-to-person route or indirectly through environmental surfaces or food, contaminated fomites and inanimate surfaces are important vehicles for the spread of the virus during norovirus outbreaks.
We developed and evaluated a protocol using macrofoam swabs for the detection and typing of human noroviruses from hard surfaces. Compared with fiber-tipped swabs or antistatic wipes, macrofoam swabs allow virus recovery (range 1.2-33.6%) from toilet seat surfaces of up to 700 cm2. The protocol includes steps for the extraction of the virus from the swabs and further concentration of the viral RNA using spin columns. In total, 127 (58.5%) of 217 swab samples that had been collected from surfaces in cruise ships and long-term care facilities where norovirus gastroenteritis had been reported tested positive for GII norovirus by RT-qPCR. Of these 29 (22.8%) could be successfully genotyped. In conclusion, detection of norovirus on environmental surfaces using the protocol we developed may assist in determining the level of environmental contamination during outbreaks as well as detection of virus when clinical samples are not available; it may also facilitate monitoring of effectiveness of remediation strategies.

A macrofoam based sampling methodology was developed and evaluated for the detection and quantification of norovirus on environmental hard surfaces.

表面上のヒトノロウイルスの検出のためのスワブサンプリング方法

Human noroviruses are a leading cause of epidemic and sporadic gastroenteritis worldwide. Because most infections are either spread directly via the ...

免疫・感染学

Prion Safety Laboratory Swipe Test

Transmission of iatrogenic prion disease has occurred from contaminated neurosurgical tools, transplant materials, and occupational exposure to prion-contaminated laboratory tools. Prions cause disease by the templated misfolding of the normal cellular form of the prion protein, PrPC, into the misfolded and pathogenic form PrPSc and are invariably fatal. Reducing iatrogenic and occupational prion transmission is challenging. First, prions can bind to and persist on surfaces for long periods of time. Second, prions are highly resistant to inactivation. Given this, surfaces can retain infectivity for long periods of time following ineffective decontamination. Not only can this pose a potential occupational risk for prion laboratory workers, but it could potentially cross-contaminate laboratory experiments utilizing sensitive prion amplification techniques. The protocol described here for a prion safety laboratory swipe test includes steps for the identification and documentation of high-traffic laboratory areas, recommended swabbing controls to ensure the validity of results, steps to identify proper responses to positive surface swabbing sites, representative results from prion swipe testing, as well as potential artifactual results. Overall, the prion safety laboratory swipe test can be implemented as part of a broader prion safety program to assess decontamination of surfaces, monitor common spaces for prion contamination, and implement the documentation of prion decontamination status.

A method to assess commonly used areas in laboratory settings for prion contamination and effective decontamination is lacking. The protocol described here provides key fundamentals for implementing a laboratory prion safety swipe test that can easily be modified to meet the individual needs of specific laboratories.

プリオン安全研究所 スワイプ試験

Transmission of iatrogenic prion disease has occurred from contaminated neurosurgical tools, transplant materials, and occupational exposure to ...

Swabbing the Urban Environment - A Pipeline for Sampling and Detection of SARS-CoV-2 From Environmental Reservoirs

To control community transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the 2020 global pandemic, most countries implemented strategies based on direct human testing, face covering, and surface disinfection. Under the assumption that the main route of transmission includes aerosols and respiratory droplets, efforts to detect SARS-CoV-2 in fomites have focused on locations suspected of high prevalence (e.g., hospital wards, cruise ships, and mass transportation systems). To investigate the presence of SARS-CoV-2 on surfaces in the urban environment that are rarely cleaned and&#160;seldomly disinfected, 350 citizens were enlisted from the greater San Diego County. In total, these citizen scientists collected 4,080 samples. An online platform was developed to monitor sampling kit delivery and pickup, as well as to collect sample data. The sampling kits were mostly built from supplies available in pandemic-stressed stores. Samples were processed using reagents that were easy to access despite the recurrent supply shortage. The methods used were highly sensitive and resistant to inhibitors that are commonly present in environmental samples. The proposed experimental design and processing methods were successful at engaging numerous citizen scientists who effectively gathered samples from diverse surface areas. The workflow and methods described here are relevant to survey the urban environment for other viruses, which are of public health concern and pose a threat for&#160;future pandemics.

A citizen science project was designed to recruit San Diego residents to collect environmental samples for SARS-CoV-2. A multilingual web-based platform was created for data submission using a user-friendly mobile device interface. A laboratory information management system facilitated the collection of thousands of geographically diverse samples with real-time outcome tracking.

都市環境のスワッピング - 環境貯留層からのSARS-CoV-2のサンプリングと検出のためのパイプライン

To control community transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the 2020 global pandemic, most countries ...

生物学

ナノハーブコーティング生地の抗菌効果

An Antimicrobial Fabric Using Nano-Herbal Encapsulation of Essential Oils

Lab coats are widely used in biohazard laboratories and healthcare facilities as protective garments to prevent direct exposure to pathogens, spills, and burns. These cotton-based protective coats provide ideal conditions for microbial growth and attachment sites due to their porous nature, moisture-holding capacity, and retention of warmth from the user&#39;s body. Several studies have demonstrated the survival of pathogenic bacteria on hospital garments and lab coats, acting as vectors of microbial transmission.
A common approach to fix these problems is the application of antimicrobial agents in textile finishing, but concerns have been raised due to the toxicity and environmental effects of many synthetic chemicals. The ongoing pandemic has also opened a window for the investigation of effective antimicrobials and eco-friendly and toxic-free formulations. This study uses two natural bioactive compounds, carvacrol and thymol, encapsulated in chitosan nanoparticles, which guarantee effective protection against four human pathogens with up to a 4-log reduction (99.99%). These pathogens are frequently detected in lab coats used in biohazard laboratories.
The treated fabrics also resisted up to 10 wash cycles with 90% microbial reduction, which is sufficient for the intended use. We made modifications to the existing standard fabric tests to better represent the typical scenarios of lab coat usage. These refinements allow for a more accurate evaluation of the effectiveness of antimicrobial lab coats and for the simulation of the fate of any accidental microbial spills that must be neutralized within a short time. Further studies are recommended to investigate the accumulation of pathogens over time on antimicrobial lab coats compared to regular protective coats.

著者スポットライト:エッセンシャルオイルのナノハーブカプセル化を使用した抗菌生地  

Antimicrobial lab coats prevent the cross-contamination of pathogen accumulation and accidental bio-spills. Here, we describe the protocol for developing a skin-friendly antimicrobial fabric using nano-herbal encapsulation and modified standard tests to precisely evaluate the efficacy and suitability for typical usage of the lab coat.

エッセンシャルオイルのナノハーブカプセル化を使用した抗菌ファブリック

Lab coats are widely used in biohazard laboratories and healthcare facilities as protective garments to prevent direct exposure to pathogens, spills, and ...

Cellular Impedance-Based Survival Kinetics Assay to Assess Virus Survival

Source: Labadie, T. et al., <a href="https://app-jove-com.remotexs.ntu.edu.sg/v/61133">Monitoring Influenza Virus Survival Outside the Host Using Real-Time Cell Analysis.</a> J. Vis. Exp. (2021).
This video describes a cellular impedance-based technique to study the survival kinetics of viruses that induce cytopathic effects. The method measures the electrical impedance of virus-infected adherent cells pre-treated with saline.

ウイルスの生存を評価するための細胞インピーダンスベースの生存速度論アッセイ

Source: Labadie, T. et al., Monitoring Influenza Virus Survival Outside the Host Using Real-Time Cell Analysis. J. Vis. Exp. (2021).
This video describes ...

JoVE Encyclopedia of Experiments

生物学的手法

アッセイ技術

細胞ベースのアッセイ

A Deferred Growth Inhibition Assay to Evaluate the Effect of Bacteria-Derived Antimicrobial Compounds

Source: Moran, J. C. et al., <a href="https://app-jove-com.remotexs.ntu.edu.sg/v/54437">Deferred Growth Inhibition Assay to Quantify the Effect of Bacteria-derived Antimicrobials on Competition.</a> J. Vis. Exp.&#160;(2016)
This video showcases the deferred growth inhibition assay, used to investigate how antimicrobial compounds produced by one type of bacteria affect another competing bacterial strain.

細菌由来の抗菌化合物の効果を評価するための遅延増殖阻害アッセイ

The following method is adapted&#160;to test competitor strains with inhibitor-producing strains.
NOTE: This protocol involves aerosol generation with ...

発生する感染性病原体の除去のための手洗いの有効性を試験する方法

要約

さらに動画を探す

この動画の章

エンテロウイルスと文化およびRT-QPCRによる水中のノロウイルス発生のEPAメソッド1615測定。ウイルスサンプルのI.コレクション

温水洗浄のウイルス消毒効果の決定

表面上のヒトノロウイルスの検出のためのスワブサンプリング方法

プリオン安全研究所スワイプ試験

都市環境のスワッピング - 環境貯留層からのSARS-CoV-2のサンプリングと検出のためのパイプライン

エッセンシャルオイルのナノハーブカプセル化を使用した抗菌ファブリック

エッセンシャルオイルのナノハーブカプセル化を使用した抗菌ファブリック

エッセンシャルオイルのナノハーブカプセル化を使用した抗菌ファブリック

ウイルスの生存を評価するための細胞インピーダンスベースの生存速度論アッセイ

細菌由来の抗菌化合物の効果を評価するための遅延増殖阻害アッセイ

発生する感染性病原体の除去のための手洗いの有効性を試験する方法

要約

さらに動画を探す

この動画の章

エンテロウイルスと文化およびRT-QPCRによる水中のノロウイルス発生のEPAメソッド1615測定。ウイルスサンプルのI.コレクション

温水洗浄のウイルス消毒効果の決定

表面上のヒトノロウイルスの検出のためのスワブサンプリング方法

プリオン安全研究所 スワイプ試験

都市環境のスワッピング - 環境貯留層からのSARS-CoV-2のサンプリングと検出のためのパイプライン

エッセンシャルオイルのナノハーブカプセル化を使用した抗菌ファブリック

エッセンシャルオイルのナノハーブカプセル化を使用した抗菌ファブリック

エッセンシャルオイルのナノハーブカプセル化を使用した抗菌ファブリック

ウイルスの生存を評価するための細胞インピーダンスベースの生存速度論アッセイ

細菌由来の抗菌化合物の効果を評価するための遅延増殖阻害アッセイ

プリオン安全研究所スワイプ試験