Name: a method to test the efficacy of handwashing for the removal of emerging infectious pathogens
Uploaded: 2017-06-07T14:00:00
Description: Watch this Scientific Journal Video about A Method to Test the Efficacy of Handwashing for the Removal of Emerging Infectious Pathogens at JoVE.com

Ce travail a été soutenu par l&#39;Agence des États-Unis pour le développement international, Office of Foreign Disaster Assistance (AID-OFDA-A-15-00026). Marlene Wolfe a été soutenue par la National Science Foundation (subvention 0966093).

Déclaration d&#39;éthique: L&#39;étude décrite ici (sur Phi6 et E. coli en tant que substitut à Ebola) a été approuvée par le Conseil d&#39;examen institutionnel du Tufts Medical Center et du campus des sciences de la santé de Tufts (no 12018); L&#39;Université de Harvard a cédé son avis au Conseil d&#39;examen institutionnel de Tufts. REMARQUE: Avant de commencer ce protocole, deux étapes doivent être complétées. Tout d&#39;abord, une version de substitution de nive...

<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. -. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Health+care+workers'+hand+contamination+levels+and+antibacterial+efficacy+of+different+hand+hygiene+methods+used+in+a+Vietnamese+hospital.">Health care workers' hand contamination levels and antibacterial efficacy of different hand hygiene methods used in a Vietnamese hospital.</a> Am J Infect Control. 42 (2), 178-181 (2014).</li><li>Steinmann, J., Nehrkorn, R., Meyer, A., Becker, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+in-vivo+protocols+for+testing+virucidal+efficacy+of+handwashing+and+hand+disinfection.">Two in-vivo protocols for testing virucidal efficacy of handwashing and hand disinfection.</a> Int J Hyg Environ Health. 196 (5), 425-436 (1995).</li><li>Weber, D. J., Sickbert-Bennett, E., Gergen, M. F., Rutala, W. A. <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+selected+hand+hygiene+agents+used+to+remove+Bacillus+atrophaeus+(a+surrogate+of+Bacillus+anthracis)+from+contaminated+hands.">Efficacy of selected hand hygiene agents used to remove Bacillus atrophaeus (a surrogate of Bacillus anthracis) from contaminated hands.</a> JAMA. 289 (10), 1274-1277 (2003).</li><li>Grayson, M. L., Melvani, S., 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=Efficacy+of+Soap+and+Water+and+Alcohol-Based+Hand-Rub+Preparations+against+Live+H1N1+Influenza+Virus+on+the+Hands+of+Human+Volunteers.">Efficacy of Soap and Water and Alcohol-Based Hand-Rub Preparations against Live H1N1 Influenza Virus on the Hands of Human Volunteers.</a> Clin Infect Dis. 48 (3), 285-291 (2009).</li><li>Oughton, M. T., Loo, V. G., Dendukuri, N., Fenn, S., Libman, M. 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+hygiene+with+soap+and+water+is+superior+to+alcohol+rub+and+antiseptic+wipes+for+removal+of+Clostridium+difficile.">Hand hygiene with soap and water is superior to alcohol rub and antiseptic wipes for removal of Clostridium difficile.</a> Infect Control Hosp Epidemiol. 30 (10), 939-944 (2009).</li><li>Liu, P., Yuen, Y., Hsiao, H. -. M., Jaykus, L. -. A., Moe, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effectiveness+of+liquid+soap+and+hand+sanitizer+against+Norwalk+virus+on+contaminated+hands.">Effectiveness of liquid soap and hand sanitizer against Norwalk virus on contaminated hands.</a> Appl Environ Micro. 76 (2), 394-399 (2010).</li><li>Savolainen-Kopra, C., Korpela, T., 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=Single+treatment+with+ethanol+hand+rub+is+ineffective+against+human+rhinovirus--hand+washing+with+soap+and+water+removes+the+virus+efficiently.">Single treatment with ethanol hand rub is ineffective against human rhinovirus--hand washing with soap and water removes the virus efficiently.</a> J Med Virol. 84 (3), 543-547 (2012).</li><li>Tuladhar, E., Hazeleger, W. C., Koopmans, M., Zwietering, M. 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. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disinfection+of+hands:+removal+of+transient+organisms.">Disinfection of hands: removal of transient organisms.</a> BMJ. 2 (5403), 230-233 (1964).</li><li>Edmonds, S. L., Zapka, C., 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=Effectiveness+of+Hand+Hygiene+for+Removal+of+Clostridium+difficile+Spores+from+Hands.">Effectiveness of Hand Hygiene for Removal of Clostridium difficile Spores from Hands.</a> Infect Control Hosp Epidemiol. 34 (3), 302-305 (2013).</li><li>Rotter, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=150+years+of+hand+disinfection-Semmelweis'+heritage.">150 years of hand disinfection-Semmelweis' heritage.</a> Hyg Med. (22), 332-339 (1997).</li><li>Hitomi, S., Baba, S., Yano, H., Morisawa, Y., Kimura, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antimicrobial+effects+of+electrolytic+products+of+sodium+chloride--comparative+evaluation+with+sodium+hypochlorite+solution+and+efficacy+in+handwashing.">Antimicrobial effects of electrolytic products of sodium chloride--comparative evaluation with sodium hypochlorite solution and efficacy in handwashing.</a> Kansensh&#333;gaku Zasshi. 72 (11), 1176-1181 (1998).</li><li>. Standard E1174-13. Standard Test Method for Evaluation of the Effectiveness of Health Care Personnel Handwash Formulations Available from: <a target="_blank" href="https://www.astm.org/">https://www.astm.org/</a> (2013)</li><li>Casanova, L. M., Weaver, S. 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. D., Agner, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+hand+eczema+severity+index+(HECSI):+A+scoring+system+for+clinical+assessment+of+hand+eczema.+A+study+of+inter-+and+intraobserver+reliability.">The hand eczema severity index (HECSI): A scoring system for clinical assessment of hand eczema. A study of inter- and intraobserver reliability.</a> Br J Dermatol. 152 (2), 302-307 (2005).</li><li>. Method 1604: Total Coliforms and Escherichia coli in Water by Membrane Filtration Using a Simultaneous Detection Technique (MI Medium) Available from: <a target="_blank" href="https://www.epa.gov/sites/production/files/2015-08/documents/method_1604_2002.pdf">https://www.epa.gov/sites/production/files/2015-08/documents/method_1604_2002.pdf</a> (2002)</li><li>Adams, M. H., Anderson, E. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Bacteriophages. , (1959).</li><li>Kao, L. S., Green, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+Variance:+Is+There+a+Difference+in+Means+and+What+Does+It+Mean?.">Analysis of Variance: Is There a Difference in Means and What Does It Mean?.</a> The Journal of surgical research. 144 (1), 158-170 (2008).</li><li>Schutz, R. W., Gessaroli, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Analysis+of+Repeated+Measures+Designs+Involving+Multiple+Dependent+Variables.">The Analysis of Repeated Measures Designs Involving Multiple Dependent Variables.</a> Research Quarterly for Exercise and Sport. 58 (2), 132-149 (1987).</li><li>Woolwine, J. D., Gerberding, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+testing+method+on+apparent+activities+of+antiviral+disinfectants+and+antiseptics.">Effect of testing method on apparent activities of antiviral disinfectants and antiseptics.</a> Antimicrob Agents Chemother. 39 (4), 921-923 (1995).</li></ol>

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...

Le lavage des mains est largement recommandé pour prévenir la propagation de la maladie, en particulier celles transmises par voie fécale-orale ou aérienne, y compris les maladies diarrhéiques et respiratoires 1 . De manière surprenante, il existe peu de preuves comparables sur l&#39;efficacité des méthodes de lavage des mains, telles que le lavage des mains avec du savon et de l&#39;eau (HWWS) et un désinfectant pour les mains à base d&#39;alcool (ABHS), sur l&#39;élimination des organismes des mains. La recherche initiale a révélé que l&#39;action mécanique du lavage des mains, par opposition à la méthode de lavage des mains, peut être responsable de la suppression de l&#39;organisme 2 , 3 . En outre, il existe peu de preuves comparatives sur lesquelles la méthode de lavage des mains est la plus efficace. Dans une revue de la littérature informelle, 14 études qui ont comparé l&#39;efficacité du savon et du désinfectant pour les mains sur l&#39;élimination des organismes ont été identifiées. Parmi ces études, cinq ont révélé que l&#39;ABHS était plus efficace 4 , </Sup&gt; 5 , 6 , 7 , 8 , sept ont révélé que le HWWS était plus efficace 9 , 10 , 11 , 12 , 13 , 14 , 15 et deux ne trouvaient aucune différence significative entre les méthodes 16 , 17 . Ces résultats sont incohérents et ne traitent pas du risque continu de maladie de la persistance d&#39;organismes dans l&#39;eau de rinçage après le lavage des mains. Dans l&#39;ensemble, les données probantes sur l&#39;efficacité comparative des méthodes de lavage des mains pour l&#39;élimination des agents pathogènes pathogènes infectieux sont limitées. Cette preuve limitée a conduit à l&#39;incertitude quant aux méthodes les plus appropriées dans les paramètres de l&#39;épidémie. Par exemple, Lors de l&#39;épidémie de maladie du virus de l&#39;Ebola (EVD) en Afrique de l&#39;Ouest de 2013 à 2016, plusieurs grands intervenants internationaux ont fourni des recommandations contradictoires pour HWWS, ABHS ou 0,05% de solutions de chlore. Médecins Sans Frontières (MSF) recommande l&#39;utilisation de solution de chlore à 0,05% pour le lavage des mains, tandis que l&#39;Organisation mondiale de la santé (OMS) recommande HWWS ou ABHS (si les mains ne sont pas visiblement souillées). L&#39;OMS va jusqu&#39;à affirmer que le chlore ne doit pas être utilisé à moins d&#39;autres options disponibles, car il est moins efficace que d&#39;autres méthodes en raison de la demande de chlore exercée par la peau 18 , 19 , 20 , 21 , 22 . De plus, les solutions de chlore sont généralement produites à partir de quatre composés de chlore différents, y compris l&#39;hypochlorite à haute teneur (HTH), l&#39;hypochlorite de sodium stabilisé (NaOCl) localisé et stabilisé et le gazonIum dichloroisocyanurate (NaDCC). Un examen systématique commandé par l&#39;OMS en réponse à l&#39;épidémie EVD en Afrique de l&#39;Ouest n&#39;a trouvé que quatre études portant sur l&#39;efficacité comparative du lavage des mains au chlore 23 . Ces études ont également produit des résultats contradictoires, et aucune de ces études n&#39;a utilisé la concentration recommandée de chlore de 0,05% pour le lavage des mains ou les microorganismes étudiés similaires au virus Ebola 10 , 24 , 25 , 26 , 27 . Ainsi, les recommandations n&#39;étaient pas fondées sur des données probantes, et il n&#39;était pas clair quelles recommandations étaient les plus efficaces. Des recherches supplémentaires sont nécessaires pour comparer les méthodes de lavage des mains pour prévenir la propagation des agents pathogènes infectieux, car les interventions de lavage des mains constituent un outil important pour prévenir la transmission des maladies épidémiques. Ces hLes recommandations sur le lavage doivent être fondées sur des preuves. Ainsi, une méthode pour tester l&#39;efficacité de la lavage des mains et la persistance de l&#39;eau de rinçage, réalisée avec des substituts ou des agents pathogènes non infectieux, a été développée 2 , 28 , 29 . Les résultats de l&#39;échantillon, en utilisant Phi6 comme substitut du virus Ebola et en utilisant Escherichia coli comme organisme témoin commun, sont présentés ici. Dans ce protocole, l&#39;efficacité de la lavage des mains et les tests de persistance de l&#39;eau de rinçage sont présentés.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Soap bar</td>
			<td>Dove</td>
			<td colspan="2">White Beauty Bar soap</td>
		</tr>
		<tr>
			<td>Alcohol-based hand sanitizer</td>
			<td>Purell</td>
			<td colspan="2">Advanced Instant Hand Sanitizer with 70% Ethyl Alcohol</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 &#181;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>E. coli</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 (47mm, 0.45 &#181;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 (47mm)</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

Le lavage des mains est largement recommandé pour prévenir la transmission des maladies infectieuses. Cependant, il existe peu de preuves comparables sur l&#39;efficacité des méthodes de lavage des mains en général. En outre, peu de preuves existent en comparant les méthodes de lavage des mains pour déterminer celles qui sont les plus efficaces pour éliminer les agents pathogènes infectieux. Des recherches sont nécessaires pour fournir des preuves des différentes approches du lavage des mains qui peuvent être utilisées pendant les épidémies de maladies infectieuses. Ici, une méthode de laboratoire pour évaluer l&#39;efficacité des méthodes de lavage des mains à l&#39;élimination des microorganismes des mains et leur persistance dans l&#39;eau de rinçage est décrite. Les mains des bénévoles sont d&#39;abord enrichies avec l&#39;organisme d&#39;essai et ensuite lavées avec chaque méthode d&#39;intérêt pour la lavage des mains. Généralement, les microorganismes de substitution sont utilisés pour protéger les sujets humains contre la maladie. Le nombre d&#39;organismes restant sur les mains des volontaires après le lavage est testé à l&#39;aide d&#39;une méthode modifiée de «jus de gant»: les mains sont placées dans des gants avec un éluEt sont nettoyés pour suspendre les microorganismes et les rendre disponibles pour analyse par filtration sur membrane (bactéries) ou en test de plaque (virus / bactériophages). L&#39;eau de rinçage produite à partir du lavage des mains est directement collectée pour analyse. L&#39;efficacité de la lavage des mains est quantifiée en comparant la valeur de réduction logarithmique entre les échantillons prélevés après le lavage des mains aux échantillons sans lavage des mains. La persistance de l&#39;eau de rinçage est quantifiée en comparant les échantillons d&#39;eau de rinçage de différentes méthodes de lavage des mains aux échantillons prélevés après le lavage des mains avec de l&#39;eau juste. Bien que cette méthode soit limitée par la nécessité d&#39;utiliser des organismes de substitution pour préserver la sécurité des volontaires humains, elle capture des aspects du lavage des mains qui sont difficiles à reproduire dans une étude in vitro et remplit les lacunes de recherche sur l&#39;efficacité du lavage des mains et la persistance des organismes infectieux au rinçage eau.

Ici, le protocole ( Figure 1 ) a été complété avec 18 volontaires, qui ont chacun été testés en utilisant à la fois E. coli et Phi6. Des différences importantes ont été trouvées entre les résultats de lavage des mains avec E. coli avec et sans charge de sol et Phi6 avec la charge du sol ( Figure 2 et Figure 3 ). Pour E. coli sans charge du sol, le lavage des m...

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

Le lavage des mains est largement recommandé pour prévenir la transmission des maladies infectieuses. Cependant, il existe peu de preuves sur lesquelles les méthodes de lavage des mains sont les plus efficaces pour éliminer les agents pathogènes des maladies infectieuses. Nous avons développé une méthode pour évaluer l&#39;efficacité des méthodes de lavage des mains lors de l&#39;élimination des microorganismes.

Une méthode pour tester l&#39;efficacité du lavage des mains pour l&#39;élimination des pathogènes infectieux émergents

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

The overall goal of this experiment is to test the comparative efficacy of handwashing options on hands that have been experimentally inoculated with an organism of interest. This method can provide information on key questions to help prevent of infectious diseases. By allowing researchers to investigate the efficacy of handwashing for particular organisms of interest.The main advantage of this technique is that it is flexible and can be adapted to allow for investigation of many different organisms. We first had the idea for this method when we realized chlorine solutions were being used for handwashing in Ebola contexts and there was no evidence for the efficacy of this at the time. After recruiting eligible subjects and preparing handwashing solutions according to the text protocol, prepare bacteria by streaking a non-pathogenic strand of E.Coli onto LB agar plates and incubate the cultures at 37 degrees celsius for 24 hours to obtain single colonies.One day before the start of the experiment, use a sterile lube to grab a single colony from the plate and inoculate 10 milliliters of LB broth. Incubate the cultures at 37 degree celsius, with shaking, overnight. On the morning of the experiment, add one milliliter of the overnight culture to 20 milliliters of fresh LB broth.Incubate the cultures for approximately two and half hours to achieve a salt density greater than 10 to the eighth CFU per milliliter. Then, use a spectrophotometer to estimate the concentration of the culture. Immediately prior to testing, prepare an inoculum composed of 68%bacteria or viral suspension and 32%sodium chloride or soil load.Swirl or gently vortex to mix. After preparing the volunteers according to the text protocol, test the pH of each volunteer's skin by placing a flat tip skin pH probe in the webbed space between the pointer and the middle fingers. Ensure that the electrode is flat against the skin and record the pH reading.Next, have the volunteer cup both hands together. Then, carefully pipette 750 microliters of the inoculate into each palm to spike the hands. When spiking volunteers'hands, it is important none of the inoculum is spilled.Remind volunteers this is important and give them the opportunity to practice with water before beginning the procedure. Have the volunteers gently rub their hands together until all surfaces of the hand are coded with the inoculate, while subjecting the hands to as little friction as possible. Instruct the volunteers to hold their hands still and away from their body for an additional 30 seconds to allow the inoculate to dry.Then wash the hands using one of the methods described in the text protocol. For hand washing with soap, use 10 milliliters of Ultrapure water to wet the hands. Have the volunteers lather their hands with soap and then rub their hands together for an additional 20 seconds.Rinse the volunteer's hands by pouring 500 milliliters of Ultrapure water at room temperature through a funnel at a flow rate of 1.5 liters per minute into a large sample collection bag and process the sample within two hours. For all chlorine solutions, pour 200 milliliters of chlorine solution over the volunteer's hands through a funnel at a flow rate of 1.5 liters per minute and have the volunteers rub their hands thoroughly. As another hand washing method, have the volunteers use hand sanitizer to wash their hands.It's important all surfaces of the hand are gently rubbed, instruct participants to focus on each part, palms, fingernails, in between fingers as they rub and then ensure the entire hand has been rinsed. For control A, after inoculation, do not perform a handwashing step. For control B, use only Ultrapure water at room temperature to perform handwashing through a funnel with a known flow rate.After handwashing, immediately place each volunteer's hand selected for testing, into a sample bag containing 75 milliliters of Leeuwin up to the wrist. Hold the top of the bag tightly around the wrist. Have the volunteers gently rub their hand in the solution for 30 seconds, taking care to reach in between the fingers and underneath the fingernails.Gently massage the hand from outside the bag for 30 seconds to ensure that the entire hand is thoroughly rinsed in the Leeuwin, all the way up to the wrist. Seal the bag and process it according to the appropriate essay. Before repeating the process with each handwashing method, have the volunteers use soap in warm water to thoroughly wash their hands in the sink.Finally, use 70%ethanol to spray the volunteer's hands until they are coated on both sides, and allow them to dry. To analyze the samples, set up a flame and a filtration manifold with sterile filtration funnels and a vacuum connection. Sterilize forceps by dipping them in ethanol and flaming them.Use the forceps to place a 0.45 micron filter onto the filtration manifold with the grid facing up. Then use a small amount of sterile PBS to wet the filter. Place the funnel on the base and pipette or pour the sample directly onto the filter.Engage the vacuum until the entire sample has passed through the membrane. Then, use sterile PBS to rinse the sides of the funnel and engage the vacuum again. Next, remove the funnel, flame sterilize the forceps and lift the filter from the base.Then place the filter gently on the medium in a petri dish with the grid facing up, ensuring that the filter lies flush against the surface. Invert the plates and incubate at 37 degrees celsius for 24 hours. Following the incubation, count the E.Coli colonies and record the data.Carry out data analysis according to the text protocol. The protocol described in this video was completed with 18 volunteers tested with E.Coli and Phi6, for E.Coli without soil load, handwashing with HTH, NaDCC and stabilized NaOCl, all resulted in significantly greater log reductions than handwashing with water only. With soil load, HTH resulted in a significantly greater log reduction of E.Coli than water only handwashing with soap and ABHS.There was no significant difference between methods for Phi6 without soil load. However, for Phi6 with soil load, water alone resulted in a greater log reduction than ABHS or stabilized NaOCl, and handwashing with soap in a greater log reduction than ABHS, stabilized NaOCl and generated NaOCl. HTH also had a greater log reduction than ABHS and stabilized NaOCl and NaDCC resulted in a greater log reduction than stabilized NaOCl and ABHS.As demonstrated in these graphs, chlorine resulted in a significantly greater log reduction of E.Coli persisting in the rinse water than handwashing with soap. Finally, the same pattern was found in Phi6 without soil load with all chlorine solutions resulting in a significantly greater reduction of Phi6 in rinse water than handwashing with soap. There were no significant decreases in rinse water with Phi6 and soil load.While attempting this procedure, it's important to remember efficacy results for each condition are calculated by comparing the results for the two controls. This means it is very important to perform the steps consistently for each handwashing option.

Research

JoVE Journal

Immunology and Infection

A 96 Well Microtiter Plate-based Method for Monitoring Formation and Antifungal Susceptibility Testing of Candida albicans Biofilms

A 96 Well Microtiter Plate-based Method for Monitoring Formation and Antifungal Susceptibility Testing of Candida albicans Biofilms

Une plaque de microtitrage à 96 puits à base Méthode de formation suivi et les tests de sensibilité antifongique des Candida albicans Les biofilms

Candida albicans remains the most frequent cause of fungal infections in an expanding population of compromised patients and candidiasis is now the third ...

Recherche

Immunologie et infection

Isolation of Brain and Spinal Cord Mononuclear Cells Using Percoll Gradients

Isolement du cerveau et les cellules mononucléées de la moelle épinière utilisant gradients de Percoll

Isolation of immune cells that infiltrate the central nervous system (CNS) during infection, trauma, autoimmunity or neurodegeneration, is often required ...

Using a Pan-Viral Microarray Assay (Virochip) to Screen Clinical Samples for Viral Pathogens

Using a Pan-Viral Microarray Assay Virochip to Screen Clinical Samples for Viral Pathogens

En utilisant un dosage Microarray pan-virale (Virochip) pour cribler des échantillons cliniques pour les pathogènes viraux

The diagnosis of viral causes of many infectious diseases is difficult due to the inherent sequence diversity of viruses as well as the ongoing emergence ...

'Bioluminescent' Reporter Phage for the Detection of Category A Bacterial Pathogens

Phage »Bioluminescent&quot; Reporter pour la détection des bactéries pathogènes de la catégorie A

Yersinia pestis and Bacillus anthracis are Category A bacterial pathogens that are the causative agents of the plague and anthrax, respectively 1. ...

Diagnosing Pulmonary Tuberculosis with the Xpert MTB/RIF Test

Diagnostic de la tuberculose pulmonaire avec le Xpert MTB / RIF test

Tuberculosis (TB) due to Mycobacterium tuberculosis (MTB) remains a major public health issue: the infection affects up to one third of the world ...

Use of Artificial Sputum Medium to Test Antibiotic Efficacy Against Pseudomonas aeruginosa in Conditions More Relevant to the Cystic Fibrosis Lung

Use of Artificial Sputum Medium to Test Antibiotic Efficacy Against Pseudomonas aeruginosa in Conditions More Relevant to the Cystic Fibrosis Lung

L&#39;utilisation de moyen d&#39;expectoration artificielle pour tester l&#39;efficacité des antibiotiques contre<em&gt; Pseudomonas aeruginosa</em&gt; Dans des conditions plus pertinents à la fibrose kystique du poumon

There  is growing concern about the relevance of in vitro antimicrobial  susceptibility tests when applied to isolates of P. aeruginosa from  cystic ...

Using Click Chemistry to Measure the Effect of Viral Infection on Host-Cell RNA Synthesis

Utilisation de Click Chemistry pour mesurer l&#39;effet de l&#39;infection virale sur Host-Cell Synthèse des ARN

Many RNA viruses have evolved the ability to inhibit host cell transcription as a means to circumvent cellular defenses. For the study of these viruses, ...

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection

In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection

Analyse in vitro de la réponse immunitaire cellulaire Myd88 médiation au virus du Nil occidental souche mutante Infection

An attenuated West Nile virus (WNV), a nonstructural (NS) 4B-P38G mutant, induced higher innate cytokine and T cell responses than the wild-type WNV in ...

A Rapid Strategy for the Isolation of New Faustoviruses from Environmental Samples Using Vermamoeba vermiformis

A Rapid Strategy for the Isolation of New Faustoviruses from Environmental Samples Using Vermamoeba vermiformis

Une stratégie rapide pour l&#39;isolement des nouveaux Faustoviruses d&#39;échantillons environnementaux Utilisation<em&gt; vermiformis Vermamoeba</em

The isolation of giant viruses is of great interest in this new era of virology, especially since these giant viruses are related to protists. Giant ...

A Simple Flow Cytometric Method to Measure Glucose Uptake and Glucose Transporter Expression for Monocyte Subpopulations in Whole Blood

Une cytométrie en flux simple méthode pour mesurer l&#39;absorption du glucose et de glucose Expression Transporter pour monocytes dans les sous-populations de sang total

Monocytes are innate immune cells that can be activated by pathogens and inflammation associated with certain chronic inflammatory diseases. Activation of ...