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 arbeidet ble støttet av United States Agency for International Development, Office of Foreign Disaster Assistance (AID-OFDA-A-15-00026). Marlene Wolfe ble støttet av National Science Foundation (stipend 0966093).

Etikkerklæring: Studien som er beskrevet her (på Phi6 og E. coli som surrogater for Ebola) ble godkjent av Institutional Review Board ved Tufts Medical Center og Tufts University Health Sciences Campus (# 12018); Harvard University ceded anmeldelse til Tufts Institutional Review Board. MERK: Før du begynner denne protokollen, må to trinn utføres. For det første må en Biosafety Level 1 (BSL-1) surrogat eller ikke-smittsom versjon av patogenet som skal studeres, som er trygt å...

<ol>
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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 på det sterkeste for å forhindre spredning av sykdom, særlig de som overføres av fekal-orale eller luftbårne ruter, inkludert diarre- og respiratoriske sykdommer 1 . Overraskende er det lite sammenlignbare bevis på effekten av håndvaskmetoder, for eksempel håndvask med såpe og vann (HWWS) og med alkoholbasert håndvern (ABHS), på fjerning av organismer fra hendene. Initial forskning har funnet ut at den mekaniske virkningen av håndvask, i motsetning til håndvaskmetoden, kan utgjøre mest organismefjerning 2 , 3 . I tillegg er det lite sammenlignende bevis på hvilken håndvaskmetode som er mest effektiv. I en uformell litteraturvurdering ble 14 studier som sammenlignet effektiviteten av såpe og håndrensemiddel på fjerning av organismer, identifisert. Av disse studiene fant fem ABHS å være mer effektive 4 , </Sup&gt; 5 , 6 , 7 , 8 , 7 fant HWWS å være mer effektive 9 , 10 , 11 , 12 , 13 , 14 , 15 og to fant ingen signifikant forskjell mellom metodene 16 , 17 . Disse funnene er inkonsekvente og tar ikke hensyn til den pågående sykdomsrisikoen fra vedvarende organismer i skyllvannet etter håndvask. Samlet sett er bevis på den komparative effekten av håndvask-metoder for fjerning av smittsomme sykdomsfremkallende patogener begrenset. Dette begrensede beviset har ført til usikkerhet om hvilke metoder som er mest hensiktsmessige i utbruddsinnstillinger. For eksempel, I løpet av Ebola Virus Disease (EVD) utbrudd i Vest-Afrika fra 2013 til 2016, ga flere store internasjonale respondenter motstridende anbefalinger for HWWS, ABHS eller 0,05% klorløsninger. Médecins Sans Frontières (MSF) anbefaler bruk av 0,05% klorløsning for håndvask, mens Verdens helseorganisasjon (WHO) anbefaler HWWS eller ABHS (hvis hendene ikke er synlig skitne). WHO går så langt som å anføre at klor ikke skal brukes med mindre det ikke finnes andre muligheter fordi det er mindre effektivt enn andre metoder på grunn av klor etterspørsel utført av huden 18 , 19 , 20 , 21 , 22 . I tillegg produseres kloroppløsninger vanligvis fra fire forskjellige klorforbindelser, inkludert høyt testhypokloritt (HTH), lokalt generert og stabilisert natriumhypokloritt (NaOCl), og sodIum dikloroisocyanurat (NaDCC). En systematisk gjennomgang på vegne av WHO som svar på EVD-utbruddet i Vest-Afrika fant nylig bare fire studier som undersøker den komparative effekten av håndvask med klor 23 . Disse studiene ga også motstridende resultater, og ingen av disse studiene brukte den anbefalte klorkonsentrasjonen på 0,05% for håndvask eller undersøkt mikroorganismer som ligner på Ebola-viruset 10 , 24 , 25 , 26 , 27 . Anbefalingene ble således ikke funnet å være bevisbaserte, og det var uklart hvilke anbefalinger som var mest effektive. Ytterligere undersøkelser er nødvendig for å sammenligne håndvaskende tilnærminger for å forhindre spredning av smittsomme patogener, da håndvaskingstiltak er et viktig verktøy for å forebygge epidemisk sykdomsoverføring. Disse hAndwashing anbefalinger må være basert på bevis. Således ble en metode for å teste håndvask effektivitet og skyllende vann persistens utført med surrogater eller ikke-smittsomme patogener, utviklet 2 , 28 , 29 . Prøveresultater, ved bruk av Phi6 som en surrogat for Ebola-viruset og bruk av Escherichia coli som en felles indikatororganisme, presenteres her. I denne protokollen presenteres håndvasking av effektivitet og skylling av vannholdighetstest.

<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 på det sterkeste for å forhindre overføring av smittsomme sykdommer. Imidlertid eksisterer det lite sammenlignbare bevis på effekten av håndvaskmetoder generelt. I tillegg finnes det lite bevis som sammenligner håndvaskmetoder for å bestemme hvilke som er mest effektive for å fjerne smittsomme patogener. Forskning er nødvendig for å gi bevis for de ulike tilnærmingene til håndvask som kan brukes under infeksjonssykdomsutbrudd. Her beskrives en laboratoriemetode for å vurdere effekten av håndvaskmetoder ved fjerning av mikroorganismer fra hender og deres persistens i skyllvann. Frivilliges hender blir først spiket med testorganismen og deretter vasket med hver håndvask-metode av interesse. Vanligvis brukes surrogatmikroorganismer for å beskytte mennesker mot sykdom. Antall organismer som gjenstår på frivilliges hender etter vask, testes ved hjelp av en modifisert &quot;hanske juice&quot; metode: hendene er plassert i hansker med en eluOg skrubbes for å suspendere mikroorganismer og gjøre dem tilgjengelige for analyse ved membranfiltrering (bakterier) eller plaqueanalyse (virus / bakteriofager). Skyll vann som er produsert fra håndvask, samles inn direkte for analyse. Håndvaskeffektivitet kvantifiseres ved å sammenligne loggreduksjonsverdien mellom prøver tatt etter håndvasking til prøver uten håndvasking. Skyll vannbestandighet kvantifiseres ved å sammenligne skyllvannsprøver fra forskjellige håndvaskmetoder til prøver samlet etter håndvask med bare vann. Selv om denne metoden er begrenset av behovet for å bruke surrogatorganismer for å bevare sikkerheten til humane frivillige, inntar det aspekter av håndvask som er vanskelige å replikere i en in vitro- studie og fyller forskningsgap på håndvaskingseffekt og persistensen av smittsomme organismer i skylling vann.

Her ble protokollen ( figur 1 ) fullført med 18 frivillige, som ble testet ved bruk av både E. coli og Phi6. Det ble funnet signifikante forskjeller mellom håndvaskresultatene med E. coli både med og uten jordbelastning og Phi6 med jordbelastning ( Figur 2 og Figur 3 ). For E. coli uten jordbelastning ga håndvask med HTH, NaDCC og stabilisert NaOCl alle signifikant st...

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 på det sterkeste for å forhindre overføring av smittsomme sykdommer. Det er imidlertid lite bevis på hvilke håndvaskmetoder som er mest effektive for å fjerne smittsomme sykdomspatogener. Vi utviklet en metode for å vurdere effekten av håndvask-metoder ved fjerning av mikroorganismer.

En metode for å teste effekten av håndvask for fjerning av nye smittsomme patogener

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

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