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

Dette arbejde blev støttet af De Forenede Nationers Agentur for International Udvikling, Office of Foreign Disaster Assistance (AID-OFDA-A-15-00026). Marlene Wolfe blev støttet af National Science Foundation (grant 0966093).

Etik erklæring: Studien beskrevet her (på Phi6 og E. coli som surrogater for Ebola) blev godkendt af Institutional Review Board ved Tufts Medical Center og Tufts University Health Sciences Campus (# 12018); Harvard University ceded anmeldelse til Tufts Institutional Review Board. BEMÆRK: Før du starter denne protokol, skal to trin udfyldes. For det første skal en Biosafety Level 1 (BSL-1) surrogat eller ikke-infektiøs version af det patogen, der skal studeres, der er sikkert a...

<ol>
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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. 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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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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...

Håndvask anbefales generelt for at forhindre spredning af sygdomme, især dem, der overføres af fækal-oral eller luftbåren rute, herunder diarré- og respiratoriske sygdomme 1 . Overraskende er der lidt sammenlignelige beviser for effekten af ​​håndvaskningsmetoder, såsom håndvask med sæbe og vand (HWWS) og med alkoholbaseret håndrensningsmiddel (ABHS), om fjernelse af organismer fra hænderne. Indledende forskning har fundet ud af, at den mekaniske virkning af håndvask, i modsætning til håndvaskefremgangsmåden, kan tegne sig for størstedelen af ​​organismernes fjernelse 2 , 3 . Derudover er der lidt sammenlignende beviser på, hvilken håndvaskningsmetode der er mest effektive. I en uformel litteraturvurdering blev 14 undersøgelser, der sammenlignede effektiviteten af ​​sæbe og håndrensningsmiddel til fjernelse af organismer, identificeret. Af disse undersøgelser fandt fem ABHS at være mere effektive 4 , </Sup&gt; 5 , 6 , 7 , 8 , 7 fandt HWWS at være mere effektive 9 , 10 , 11 , 12 , 13 , 14 , 15 og to fandt ingen signifikant forskel mellem fremgangsmåderne 16 , 17 . Disse resultater er inkonsekvente og tager ikke højde for den igangværende risiko for sygdom fra vedvarende organismer i skyllevandet efter håndvask. Samlet set er beviser for den komparative effektivitet af håndvaskefremgangsmåder til fjernelse af infektionssygdomsfremkaldende patogener begrænset. Dette begrænsede bevis har ført til usikkerhed om, hvilke metoder der er mest hensigtsmæssige i udbrudningsindstillinger. For eksempel, I løbet af udbruddet af Ebola Virus Disease (EVD) i Vestafrika fra 2013 til 2016 fremlagde flere store internationale respondenter modstridende anbefalinger for HWWS, ABHS eller 0,05% chloropløsninger. Médecins Sans Frontières (MSF) anbefaler brug af 0,05% chloropløsning til håndvask, mens Verdenssundhedsorganisationen (WHO) anbefaler HWWS eller ABHS (hvis hænderne ikke er synligt snavset). WHO går så langt at sige, at klor ikke bør anvendes, medmindre der ikke findes andre muligheder, fordi det er mindre effektivt end andre metoder på grund af klorens efterspørgsel udført af huden 18 , 19 , 20 , 21 , 22 . Derudover fremstilles chloropløsningerne sædvanligvis af fire forskellige chlorforbindelser, herunder højtesthypochlorit (HTH), lokalt genereret og stabiliseret natriumhypochlorit (NaOCl) og sodIum dichloroisocyanurat (NaDCC). En systematisk gennemgang af WHO som reaktion på EVD-udbruddet i Vestafrika fandt for nylig kun fire undersøgelser, der undersøgte den komparative effekt af håndvask med klor 23 . Disse undersøgelser frembragte også modstridende resultater, og ingen af ​​disse undersøgelser anvendte den anbefalede klorkoncentration på 0,05% til håndvask eller undersøgte mikroorganismer svarende til Ebola-viruset 10 , 24 , 25 , 26 , 27 . Anbefalingerne blev således ikke fundet bevisbaserede, og det var uklart hvilke anbefalinger der var mest effektive. Yderligere forskning er nødvendig for at sammenligne håndvaskefremgangsmåder for at forhindre spredning af infektiøse patogener, da håndvaskeinterventioner er et vigtigt redskab til at forebygge epidemisk sygdomsoverførsel. Disse hAndwashing anbefalinger skal være baseret på bevis. Således blev en metode til testning af håndvaskeffektivitet og skyllevandsperistens udført med surrogater eller ikke-infektiøse patogener udviklet 2 , 28 , 29 . Prøveresultater, der anvender Phi6 som en surrogat for Ebola-viruset og ved anvendelse af Escherichia coli som en fælles indikatororganisme, er præsenteret her. I denne protokol præsenteres håndvaskning af effektivitet og skylning af vandpersistensforsøg.

<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

Håndvask anbefales generelt for at forhindre overførsel af smitsomme sygdomme. Imidlertid findes der kun lidt sammenlignelige beviser for effektiviteten af ​​håndvaskningsmetoder generelt. Derudover findes der lidt beviser, der sammenligner håndvaskningsmetoder for at bestemme, hvilke der er mest effektive til fjernelse af infektiøse patogener. Forskning er nødvendig for at fremlægge bevis for de forskellige tilgange til håndvask, der kan anvendes i forbindelse med smitsomme sygdomsudbrud. Her beskrives en laboratoriemetode til vurdering af effektiviteten af ​​håndvaskefremgangsmåder til fjernelse af mikroorganismer fra hænder og deres persistens i skyllevand. Frivilliges hænder spidses først med testorganismen og vaskes derefter med hver håndvaskefremgangsmåde af interesse. Generelt anvendes surrogatmikroorganismer til at beskytte mennesker mod sygdom. Antallet af organismer, der forbliver på frivilliges hænder efter vask, testes ved hjælp af en ændret &quot;handske juice&quot; metode: Hænderne placeres i handsker med en eluOg skrubber for at suspendere mikroorganismerne og gøre dem tilgængelige til analyse ved membranfiltrering (bakterier) eller plaqueanalyse (vira / bakteriofager). Skyllevand fra håndvasken indsamles direkte til analyse. Håndvask effektivitet kvantificeres ved at sammenligne logreduktionsværdien mellem prøver taget efter håndvask til prøver uden håndvask. Skyllevandstannelsen kvantificeres ved at sammenligne skyllevandsprøver fra forskellige håndvaskemetoder til prøver opsamlet efter håndvask med bare vand. Selv om denne metode er begrænset af behovet for at anvende surrogatorganismer for at bevare sikkerheden hos mennesker, frivillige, tager det fat på aspekter af håndvask, der er vanskelige at replikere i et in vitro- studie og fylder forskningsmæssige huller i håndvaskningsevnen og persistensen af ​​infektiøse organismer i skylning vand.

Her blev protokollen ( figur 1 ) afsluttet med 18 frivillige, som hver blev testet under anvendelse af både E. coli og Phi6. Der blev fundet signifikante forskelle mellem håndvaskresultater med E. coli både med og uden jordbelastning og Phi6 med jordbelastning ( Figur 2 og Figur 3 ). For E. coli uden jordbelastning resulterede håndvask med HTH, NaDCC og stabiliseret Na...

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

Håndvask anbefales generelt for at forhindre overførsel af smitsomme sygdomme. Der er dog kun få beviser på, hvilke håndvaskningsmetoder der er mest effektive til fjernelse af infektionssygdomspatogener. Vi udviklede en metode til vurdering af effektiviteten af ​​håndvaskningsmetoder ved fjernelse af mikroorganismer.

En metode til at teste effektiviteten af ​​håndvask til fjernelse af nye smitsomme patogener

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

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En metode til at teste effektiviteten af ​​håndvask til fjernelse af nye smitsomme patogener

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