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

Questo lavoro è stato sostenuto dall&#39;Agenzia degli Stati Uniti per lo Sviluppo Internazionale, Ufficio di Assistenza Estera Del Disastro (AID-OFDA-A-15-00026). Marlene Wolfe è stata sostenuta dalla National Science Foundation (sovvenzione 0966093).

Dichiarazione di etica: Lo studio qui descritto (su Phi6 ed E. coli come surrogati per Ebola) è stato approvato dal consiglio di revisione istituzionale presso Tufts Medical Center e Tufts University Health Sciences Campus (# 12018); L&#39;Università di Harvard ha ceduto la revisione al consiglio di revisione istituzionale di Tufts. NOTA: Prima di iniziare questo protocollo, è necessario completare due passaggi. In primo luogo, deve essere identificata e selezionata una versione ...

<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...

La lavata a mano è ampiamente raccomandata per prevenire la diffusione delle malattie, in particolare quelle trasmesse dalla via fecale-orale o aerea, comprese le malattie diarrea e respiratoria 1 . Sorprendentemente, non vi è alcuna prova comparabile sull&#39;efficacia dei metodi di lavaggio a mano, come la lavaggio a mano con sapone e acqua (HWWS) e con il liquido sanitario a base di alcol (ABHS), sulla rimozione di organismi dalle mani. La ricerca iniziale ha scoperto che l&#39;azione meccanica della lavaggio a mano, al contrario del metodo di lavaggio a mano, può rappresentare la maggior parte delle rimozioni dell&#39;organismo 2 , 3 . Inoltre, non vi è alcuna prova comparativa su quale metodo di lavaggio a mano è più efficace. In una revisione informale della letteratura, sono stati identificati 14 studi che hanno confrontato l&#39;efficacia del sapone e del sanitizzatore delle mani sulla rimozione degli organismi. Di questi studi, cinque hanno trovato ABHS per essere più efficaci 4 , </Sup&gt; 5 , 6 , 7 , 8 , sette trovarono HWWS per essere più efficaci 9 , 10 , 11 , 12 , 13 , 14 , 15 e due non trovarono alcuna differenza significativa tra i metodi 16 , 17 . Questi risultati sono incoerenti e non affrontano il rischio continuo di malattia dalla persistenza degli organismi nell&#39;acqua di risciacquo dopo la lavata a mano. Nel complesso, l&#39;evidenza sull&#39;efficacia comparata dei metodi di lavaggio a mano per la rimozione di patogeni causanti malattie infettive è limitata. Questa prova limitata ha portato all&#39;incertezza circa quali metodi sono più appropriati nelle impostazioni dell&#39;epidemia. Per esempio, Durante l&#39;epidemia di virus di Ebola (EVD) in Africa occidentale dal 2013 al 2016, numerosi intervistati internazionali hanno fornito raccomandazioni contraddittorie per HWWS, ABHS o soluzioni di cloro da 0,05%. Médecins Sans Frontières (MSF) raccomanda l&#39;uso di una soluzione di cloro da 0,05% per la lavata a mano, mentre l&#39;Organizzazione Mondiale della Sanità (WHO) raccomanda HWWS o ABHS (se le mani non sono visibilmente sporche). L&#39;OMS va addirittura affermando che il cloro non deve essere utilizzato a meno che non siano disponibili altre opzioni, perché è meno efficace di altri metodi a causa della domanda di cloro esercitata dalla pelle 18 , 19 , 20 , 21 , 22 . Inoltre, le soluzioni di cloro vengono comunemente prodotte da quattro diversi composti di cloro, tra cui l&#39;ipoclorito ad alta test (HTH), l&#39;ipoclorito di sodio localizzato e stabilizzato (NaOCl) e la sodioIodicloroisocianurato (NaDCC). Una revisione sistematica commissionata dall&#39;OMS in risposta all&#39;epidemia di EVD in Africa occidentale ha recentemente trovato solo quattro studi che indagano sull&#39;efficacia comparativa della lavaggio a mano con cloro 23 . Questi studi hanno inoltre prodotto risultati conflittuali e nessuno di questi studi ha utilizzato la concentrazione raccomandata di cloro dello 0,05% per la lavata a mano o studiati microorganismi simili al virus Ebola 10 , 24 , 25 , 26 , 27 . Pertanto, le raccomandazioni non sono state ritenute basate su prove e non era chiaro quali raccomandazioni erano più efficaci. Sono necessarie ulteriori ricerche per confrontare gli approcci di lavaggio a mano per prevenire la diffusione degli agenti patogeni, poiché gli interventi di lavaggio a mano sono uno strumento importante per prevenire la trasmissione di malattie epidemiche. Questi hLe raccomandazioni di lavaggio devono essere basate su prove. Pertanto, è stato sviluppato un metodo per testare l&#39;efficacia della lavaggio e la persistenza dell&#39;acqua di risciacquo, eseguita con surrogati o agenti patogeni non infettivi, 2 , 28 e 29 . I risultati del campione, utilizzando Phi6 come surrogato per il virus Ebola e utilizzando Escherichia coli come un comune indicatore organismo, sono qui presentati. In questo protocollo viene presentata l&#39;efficacia della lavaggio e il risciacquo delle prove di persistenza dell&#39;acqua.

<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

La lavata a mano è ampiamente raccomandata per prevenire la trasmissione delle malattie infettive. Tuttavia, esistono scarse prove comparabili sull&#39;efficacia dei metodi di lavaggio a mano in generale. Inoltre, esistono poche prove che consentono di confrontare i metodi di lavaggio a mano per determinare quali sono i più efficaci per la rimozione di patogeni infettivi. È necessaria la ricerca per fornire prove sui diversi approcci di lavaggio a mano che possono essere utilizzati durante i focolai di malattie infettive. Qui viene descritto un metodo di laboratorio per valutare l&#39;efficacia dei metodi di lavaggio a mano per rimuovere i microrganismi dalle mani e la loro persistenza nell&#39;acqua di risciacquo. Le mani dei volontari sono dapprima forate con l&#39;organismo di prova e poi lavate con ogni metodo di lavaggio a mano d&#39;interesse. In generale, i microrganismi surrogati vengono utilizzati per proteggere i soggetti umani dalla malattia. Il numero di organismi che rimangono sulle mani dei volontari dopo il lavaggio viene testato utilizzando un metodo &quot;succo di guanti&quot; modificato: le mani vengono collocate in guanti con un eluE vengono sciacquati per sospendere i microrganismi e renderli disponibili per l&#39;analisi mediante filtrazione a membrana (batteri) o test di placca (virus / batteriofagi). L&#39;acqua di risciacquo prodotta dalla lavata a mano viene direttamente raccolta per l&#39;analisi. L&#39;efficacia del lavaggio manuale viene quantificata confrontando il valore di riduzione del log tra i campioni prelevati dopo lavaggio a campioni senza lavaggio a mano. La persistenza della risciacquatura dell&#39;acqua è quantificata confrontando i campioni di acqua di risciacquo da vari metodi di lavaggio a mano a campioni prelevati dopo lavaggio a mano con acqua sufficiente. Mentre questo metodo è limitato dalla necessità di utilizzare organismi surrogati per preservare la sicurezza dei volontari umani, raccoglie aspetti di lavaggio a mano che sono difficili da replicare in uno studio in vitro e riempie le lacune di ricerca sull&#39;efficacia della lavaggio e la persistenza degli organismi infettivi in ​​risciacquo acqua.

Qui, il protocollo ( Figura 1 ) è stato completato con 18 volontari, che sono stati testati sia con E. coli che Phi6. Differenze significative sono state riscontrate tra i risultati di lavaggio a mano con E. coli sia con e senza carico del suolo e Phi6 con carico del suolo ( Figura 2 e Figura 3 ). Per E. coli senza carico del suolo, la lavata a mano con HTH, NaDCC e NaOCl...

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

La lavata a mano è ampiamente raccomandata per prevenire la trasmissione delle malattie infettive. Tuttavia, non vi è alcuna prova su cui i metodi di lavaggio della mano sono più efficaci nella rimozione di patogeni delle malattie infettive. Abbiamo sviluppato un metodo per valutare l&#39;efficacia dei metodi di lavaggio a mano alla rimozione dei microrganismi.

Un metodo per testare l&#39;efficacia della lavata a mano per la rimozione di nuovi patogeni infettivi

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.

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