이 작업은 미국 국제 재난 지원 센터 (AID-OFDA-A-15-00026)에 의해 지원되었습니다. 말린 울프 (Marlene Wolfe)는 국립 과학 재단 (National Science Foundation)의 후원을 받았습니다 (부여 0966093).

<ol>
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저자는 경쟁적인 금전적 이해 관계가 없다고 선언합니다.

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

손씻기는 설사 및 호흡기 질환을 포함하여 대변 또는 구강 경로로 전염되는 질병의 확산을 막기 위해 널리 권장됩니다 1 . 놀랍게도 비누와 물 (HWWS)로 손을 닦기, 알코올 기반 손 소독제 (ABHS)로 손에서 생물을 제거하는 것과 같은 손씻기 방법의 효능에 대한 비교 가능한 증거는 거의 없다. 초기 연구에 따르면 손씻기 방법과 달리 손씻기의 기계적 작용이 대부분의 유기물 제거에 영향을 줄 수 있습니다. 또한 손씻기 방법이 가장 효과적인 비교 증거는 거의 없습니다. 비공식 문헌 검토에서 비누와 손 소독제의 효능을 유기체 제거에 비교 한 14 개의 연구가 확인되었다. 이 연구들 중 5 명은 ABHS가 더 효과적이라는 것을 발견했습니다 4 , </7 명은 HWWS가 9 , 10 , 11 , 12 , 13 , 14 , 15 로 더 효과적이라는 것을 발견하였고 2 명은 방법 16 과 17 사이에 유의 한 차이가 없음을 발견했다. 이러한 발견은 일관성이없고 손씻기 후 헹굼물에 잔류하는 유기체의 질병으로 인한 지속적인 위험을 언급하지 않습니다. 전반적으로, 감염성 질병을 유발하는 병원균 제거를위한 손씻기 방법의 비교 효능에 대한 증거는 제한적이다. 이러한 제한된 증거로 인해 어떤 방법이 질병 발생 상황에 가장 적합한 지에 대한 불확실성이 제기되었습니다. 예를 들어2013 년부터 2016 년까지 서 아프리카의 에볼라 바이러스 질병 (EVD) 발병 기간 동안 몇몇 대규모 국제 대응자가 HWWS, ABHS 또는 0.05 % 염소 솔루션에 대한 모순 된 권장 사항을 제시했습니다. 세계 보건기구 (WHO)가 HWWS 또는 ABHS (손이 눈에 보이지 않는 경우)를 권장하는 반면, Médecins Sans Frontières (MSF)는 0.05 % 염소 용액을 사용하여 손씻기를 권장합니다. WHO는 염소 요구량이 피부 18 , 19 , 20 , 21 , 22 로 인해 다른 방법보다 덜 효과적이므로 다른 옵션을 사용할 수 없다면 염소를 사용해서는 안된다는 것을 밝히고있다. 또한, 염소 용액은 일반적으로 하이 테아 차아 염소산염 (HTH), 국부적으로 생성되고 안정화 된 차아 염소산 나트륨 (NaOCl) 및 염소산 나트륨(NaDCC)이다. 서 아프리카에서의 EVD 발생에 대한 응답으로 WHO가 의뢰 한 체계적인 검토는 최근 염소로 손씻기의 비교 효능을 조사한 4 개의 연구만을 발견했다. 이 연구들은 또한 상반되는 결과를 가져 왔으며,이 연구들 중 어느 것도 에볼라 바이러스 10 , 24 , 25 , 26 , 27 과 유사한 손씻기 또는 조사 미생물에 대해 0.05 %의 권장 염소 농도를 사용하지 않았습니다. 따라서 권장 사항은 증거에 기반한 것으로 밝혀지지 않았고 어떤 권장 사항이 가장 효과적 이었는지 분명하지 않았습니다. 손씻기 중재는 전염병 전파를 예방하는 중요한 도구이기 때문에 감염 병원체의 확산을 막기위한 손씻기 접근법을 비교하기 위해서는 추가 연구가 필요합니다. 이 h세탁 권고는 증거에 근거해야합니다. 따라서 대리모 또는 비 감염성 병원체를 사용하여 실시한 손씻기 효능 및 헹굼 물 지속성을 테스트하는 방법이 개발되었습니다 2 , 28 , 29 . 에볼라 바이러스의 대리인으로 Phi6을 사용하고 일반적인 지시 생물로 대장균 을 사용하는 샘플 결과가 여기에 제시됩니다. 이 프로토콜에서는 손씻기 효능 및 헹굼 수 지속 테스트가 제공됩니다.

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

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

감염성 질병 전파를 예방하기 위해 손씻기가 널리 권장됩니다. 그러나 일반적으로 손씻기 방법의 효능에 대한 비교 가능한 증거는 거의 없다. 또한 손씻기 방법을 비교하여 감염성 병원체를 제거하는데 가장 효과적인 방법을 결정하는 증거는 거의 없다. 전염병 발생시 사용할 수있는 손씻기에 대한 다양한 접근법에 대한 증거를 제공하기위한 연구가 필요합니다. 여기에는 손에서 미생물을 제거 할 때의 손씻기 방법의 효과와 헹굼 수의 지속성을 평가하는 실험실 방법이 설명되어 있습니다. 자원 봉사자의 손에 시험 생물을 먼저 넣고 손으로 씻는 방법으로 씻어 낸다. 일반적으로 대리 미생물은 질병으로부터 사람 피험자를 보호하기 위해 사용됩니다. 세척 후 자원 봉사자의 손에 남아있는 생물의 수는 변형 된 &quot;장갑 주스&quot;방법을 사용하여 시험합니다 : 손은 elu미생물을 부유시키고 멤브레인 여과 (박테리아) 또는 플라크 분석 (바이러스 / 박테리오파지)에 의한 분석에 사용할 수 있도록 스크러빙됩니다. 손씻기에서 생성 된 헹굼 수는 분석을 위해 직접 수집됩니다. 손씻기의 효능은 손씻기 후에 채취 한 시료와 손씻기가없는 시료의 대수 감소 값을 비교하여 계량화합니다. 헹굼 물 지속성은 다양한 손 씻기 방법에서 씻어 낸 물 시료를 물로 손씻기 후에 수집 한 시료와 비교하여 계량화됩니다. 이 방법은 인간 자원 봉사자의 안전을 지키기 위해 대리 조직을 사용할 필요가 있기 때문에 제한적이지만 체외 연구에서 복제하기 어려운 손씻기의 양상을 파악하고 손씻기 효과 및 헹굼에서 감염 생물의 지속성에 대한 연구 틈을 메운다 물.

여기에서 프로토콜 ( 그림 1 )은 18 명의 자원 봉사자와 함께 완료되었으며 각각은 E.coli 와 Phi6를 사용하여 테스트되었습니다. 토양 부하가 있거나없는 대장균 과 토양 부하가있는 Phi6 ( 그림 2 및 그림 3 )과의 손 세척 결과간에 유의 한 차이가 발견되었습니다. 토양 부하가없는 대장균의 경우, HTH, NaDCC 및 안정화 된 NaOCl을 사용한 손씻기는 모두 물로 손을 씻는 것보다 훨씬 더 큰 로그 감소를 가져왔다 (F (6,102) = 2.72, p = 0.034). 토양 부하로, HTH는 물 만, HWWS 및 ABHS (F (6,102) = 3.94, p &lt;0.001)보다 대장균의 대폭 감소를 가져왔다. 토양 부하가없는 Phi6의 방법 간에는 유의 한 차이가 없었다 (F (6,66) = 2.04, p = 0.073). 그러나 토양 부하가있는 Phi6의 경우 (F (6,102) = 7.01, p &lt;0.001), 물만으로는 greABHS, 안정화 된 NaOCl 및 HWWS에 비해 로그 감소가 ABHS, 안정화 된 NaOCl 및 생성 된 NaOCl보다 큰 로그 감소로 나타났다. HTH는 또한 ABHS 및 안정화 된 NaOCl보다 더 큰 로그 감소를 가지며, NaDCC는 안정화 된 NaOCl 및 ABHS보다 큰 로그 감소를 초래 하였다. HTH는 조건들에 대해 가장 일관되게 잘 수행되었지만, 많은 신뢰 구간이 0.5 log보다 작고 1.5 log 감소보다 많은 경우, 많은 결과가 과다 해석되지 않도록주의해야합니다. 헹굼 수에서 염소는 HWWS보다 헹굼 수에서 지속되는 대장균의 대폭 감소를 가져왔다 (토양 부하가없는 경우, F (4,68) = 331.7, p &lt;0.001, 토양 부하 F (4,68) ) = 162.44, p &lt;0.001) ( 도 4 ). 이 같은 패턴은 토양 부하가없는 Phi6에서 발견되었다 ((F (4,43) = 8.95, P &lt;0.001). 모든 염소 용액은HWWS보다 헹굼 수에서 Phi6가 현저하게 더 많이 감소합니다. Phi6와 토양 부하를 가진 헹굼 수의 지속성에는 유의 한 차이가 없었다 ((F (4,67) = 3.35, p = 0.071) ( 그림 5 ). <img alt="그림 1" class="xfigimg" src="/files/ftp_upload/55604/55604fig1.jpg" /> 그림 1 : 실험 개요. 손씻기의 각 라운드마다 수행되는 5 단계는 1) pH 테스트, 2) 손 접종, 3) 손 씻기, 4) 손 씻기, 5) 테스트 된 8 가지 조건 각각에 대한 손 오염 제거 등이 있습니다. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55604/55604fig1large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 2" class="xfigimg" src="/files/ftp_upload/55604/55604fig2.jpg" /> 도표 2 : E.coli 손 세척 res<html> ults. 핸드 워싱 방법과 비교하여, 손씻기 방법은 대장균 에서 토양 부하없이 1.94-3.01 및 토양 부하로 2.18-3.34의 평균 로그 감소를 가져왔다. 물로 손을 씻음으로써 두 조건 (1.94 및 2.18 로그)에서 대장균의 감소가 가장 적다는 사실이 입증되었습니다. NaDCC로 손을 씻는 것은 토양 부하 (3.01)없이 가장 큰 감소를 가져 왔고 HTH는 토양 부하 (3.34)로 가장 큰 감소를 가져왔다. 차트에서이 선은 유기체의 퍼센트 감소를 나타내며 오차 막대는 로그 감소의 표준 오류를 나타냅니다. Ctrl B, 컨트롤 B; HWWS, 비누로 손씻기; ABHS, 알코올 기반 손 소독제; HTH, 하이포 아 염소산염 시험; NaDCC, 나트륨 디클로로이 소시 아누 레이트; st NaOCl, 안정화 된 차아 염소산 나트륨; NaOCl 생성, 차아 염소산 나트륨 생성. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55604/55604fig2large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> ve_content &quot;fo : keep-together.within-page =&quot;1 &quot;&gt; <img alt="그림 3" class="xfigimg" src="/files/ftp_upload/55604/55604fig3.jpg" /> 그림 3 : Phi6 손씻기 결과. 수동 세척을하지 않은 것에 비해 손 세정법은 Phi6에서 토양 부하가없는 경우 2.44-3.06, 토양 부하가있는 경우 2.71-3.69의 평균 로그 감소를 가져 왔습니다. 비누로 손을 씻는 것은 토양 부하 (2.44)없이 Phi6가 가장 적게 감소하고, 안정화 된 NaOCl로 손씻는 것은 토양 부하 (2.71)로 최소 감소로 나타납니다. 생성 된 NaOCl로 손을 씻음으로써 토양 부하 (3.06)없이 가장 큰 감소를 가져 왔으며 비누로 손을 씻는 것은 토양 부하 (3.69)로 가장 큰 감소를 가져 왔습니다. 차트에서이 선은 유기체의 퍼센트 감소를 나타내며 오차 막대는 로그 감소의 표준 오류를 나타냅니다. Ctrl B, 컨트롤 B; HWWS, 비누로 손씻기; ABHS, 알코올 기반 손 소독제; HTH, 하이포 아 염소산염 시험; NaDCC, 나트륨 디클로로이 소시 아누 레이트;st NaOCl, 안정화 된 차아 염소산 나트륨; NaOCl 생성, 차아 염소산 나트륨 생성. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55604/55604fig3large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 4" class="xfigimg" src="/files/ftp_upload/55604/55604fig4.jpg" /> 그림 4 : 대장균 손 헹굼 결과. 수세만으로 손을 씻을 때와 비교할 때 헹굼 수에 남아있는 대장균 의 평균 대수 감소는 토양 부하가없는 경우 0.28-4.77이었고 토양 부하가있는 경우 0.21-4.49였다. 토양 부하가 있든 없든 모두 비누로 손씻기에서 가장 작은 감소가 발견되었습니다 (0.28 및 0.21). 가장 큰 감소는 토양 부하가없는 안정화되고 생성 된 NaOCl (모두 4.77)과 HTH 및 토양 부하를 갖는 NaOCl 생성으로 관찰되었다. 도표에서, 선은 유기체의 퍼센트 감소를 나타내며, 오차 막대는로그 감소의 표준 오류. HWWS, 비누로 손씻기; ABHS, 알코올 기반 손 소독제; HTH, 하이포 아 염소산염 시험; NaDCC, 나트륨 디클로로이 소시 아누 레이트; st NaOCl, 안정화 된 차아 염소산 나트륨; NaOCl 생성, 차아 염소산 나트륨 생성. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55604/55604fig4large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 5" class="xfigimg" src="/files/ftp_upload/55604/55604fig5.jpg" /> 그림 5 : Phi6 핸드 린스 결과. 수세만으로 손을 씻을 때와 비교할 때 헹굼 수에 남아있는 Phi6의 평균 대수 감소는 토양 부하가없는 경우 1.26-2.02이었고 토양 부하가있는 경우 1.30-2.20이었다. 토양 부하의 경우, 비누로 손을 씻는 데있어 가장 작은 감소가 발견되었습니다 (1.26). 토양 부하가 없으면 HTH는 최소 감소 (2.02)를 나타냈다. 가장 큰 감소는 NaDCC로 토양 부하가 있거나없는 경우 모두 관찰되었다(2.02 및 2.20). 차트에서이 선은 유기체의 퍼센트 감소를 나타내며 오차 막대는 로그 감소의 표준 오류를 나타냅니다. HWWS, 비누로 손씻기; ABHS, 알코올 기반 손 소독제; HTH, 하이포 아 염소산염 시험; NaDCC, 나트륨 디클로로이 소시 아누 레이트; st NaOCl, 안정화 된 차아 염소산 나트륨; NaOCl 생성, 차아 염소산 나트륨 생성. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/55604/55604fig5large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a>

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

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

감염성 질병 전파를 예방하기 위해 손씻기가 널리 권장됩니다. 그러나 어떤 손씻기 방법이 전염병 병원균을 제거하는데 가장 효과적인지에 대한 증거는 거의 없다. 우리는 미생물 제거시 손씻기 방법의 효능을 평가하는 방법을 개발했습니다.

신생 전염병 병원체 제거를위한 손씻기의 효능을 시험하는 방법

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

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

Universidad San Sebastian

Research

JoVE Journal

Immunology and Infection

19.0K 조회수.  Tufts University.  감염성 질병 전파를 예방하기 위해 손씻기가 널리 권장됩니다. 그러나 어떤 손씻기 방법이 전염병 병원균을 제거하는데 가장 효과적인지에 대한 증거는 거의 없다. 우리는 미생물 제거시 손씻기 방법의 효능을 평가하는 방법을 개발했습니다. 

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

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

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

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

엔테로 바이러스 및 문화 RT-qPCR에 의해 물에서 노로 바이러스의 발생의 EPA 방법 1615 측정. 바이러스 샘플의 I. 컬렉션

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

연구

환경과학

Determining Viral Disinfection Efficacy of Hot Water Laundering

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

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

온수 세탁의 바이러스 소독 효능 결정

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

공학

Antimicrobial Synergy Testing by the Inkjet Printer-assisted Automated Checkerboard Array and the Manual Time-kill Method

As rates of multidrug-resistant (MDR) pathogens continue to rise, outpacing the development of new antimicrobials, novel approaches to treatment of MDR bacteria are increasingly becoming a necessity. One such approach is combination therapy, in which two or more antibiotics are used together to treat an infection against which one or both of the drugs may be ineffective alone. When two drugs, in combination, exert a greater than additive effect, they are considered synergistic. In vitro investigation of synergistic activity is an important first step in evaluating the possible efficacy of drug combinations. Two main in vitro synergy testing methods have been developed: the checkerboard array and the time-kill study. In this paper, we present an automated checkerboard array method that makes use of inkjet printing technology to increase the efficiency and accuracy of this technique, as well as a standard manual time-kill synergy method. The automated checkerboard array can serve as a high-throughput screening assay, while the manual time-kill study provides additional, complementary data on synergistic activity and killing.
The checkerboard array is a modification of standard minimum inhibitory concentration (MIC) testing, in which bacteria are incubated with antibiotics at different concentration combinations and evaluated for growth inhibition after overnight incubation. Manual performance of the checkerboard array requires a laborious and error-prone series of calculations and dilutions. In the automated method presented here, the calculation and dispensing of required antibiotic stock solution volumes are automated through the use of inkjet printer technology. In the time-kill synergy assay, bacteria are incubated with the antibiotics of interest, both together and individually, and sampled at intervals over the course of 24 h for quantitative culture. The results can determine whether a combination is synergistic and whether it is bactericidal, and provide data on inhibition and killing of bacteria over time.

Antimicrobial synergy testing is used to evaluate the effect of two or more antibiotics used in combination and is typically performed by one of two methods: the checkerboard array or the time-kill assay. Here, we present an automated, inkjet printer-assisted checkerboard array synergy technique and a classic time-kill synergy study.

잉크젯 프린터 기반으로 테스트 자동화 된 바둑판 배열 및 수동 시간 죽 일 방법 항균 시너지

As rates of multidrug-resistant (MDR) pathogens continue to rise, outpacing the development of new antimicrobials, novel approaches to treatment of MDR ...

면역학 및 감염병학

Prion Safety Laboratory Swipe Test

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

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

Prion 안전 연구소 스와이프 테스트

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

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

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

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

도시 환경 면봉 - 환경 저수지에서 SARS-CoV-2의 샘플링 및 검출을 위한 파이프라인

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

생물학

Evaluation of the Efficacy of Organic Peroxyacids for Eradicating Dairy Biofilms Using an Approach Combining Static and Dynamic Methods

The presence of biofilms in the dairy industry is of major concern, as they may lead to the production of unsafe and altered dairy products due to their high resistance to most clean-in-place (CIP) procedures frequently used in processing plants. Therefore, it is imperative to develop new biofilm control strategies for the dairy industry. This protocol is aimed at evaluating the efficacy of organic peroxyacids (peracetic, perpropionic, and perlactic acids and a commercial peracetic acid-based disinfectant) for eradicating dairy biofilms using a combination of static and dynamic methods. All the disinfectants were tested on the strongest biofilm-producing bacteria in either a single or a mixed biofilm using the minimum biofilm eradication concentration (MBEC) assay, a static high-throughput screening method. A contact time of 5 min with the disinfectants at the recommended concentrations successfully eradicated both the single and mixed biofilms. Studies are currently ongoing to confirm these observations using the Center for Disease Control (CDC) biofilm reactor, a dynamic method to mimic in situ conditions. This type of bioreactor enables the use of a stainless-steel surface, which constitutes most industrial equipment and surfaces. The preliminary results from the reactor appear to confirm the efficacy of organic peroxyacids against biofilms. The combined approach described in this study may be used to develop and test new biological or chemical formulations for controlling biofilms and eradicating microorganisms.

This protocol describes an approach combining static and dynamic methods to evaluate the efficacy of organic peroxyacids for eradicating biofilms in the dairy industry. This approach may also be used to test the effectiveness of new biological or chemical formulations for controlling biofilms.

정적 방법과 동적 방법을 결합한 접근법을 사용하여 낙농 생물막을 박멸하기 위한 유기 과산화산의 효능 평가

The presence of biofilms in the dairy industry is of major concern, as they may lead to the production of unsafe and altered dairy products due to their ...

Nano-Herbal-Coated Fabrics의 항균 효능

An Antimicrobial Fabric Using Nano-Herbal Encapsulation of Essential Oils

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

저자 스포트라이트: 에센셜 오일의 나노 허브 캡슐화를 사용한 항균 직물  

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

에센셜 오일의 나노 허브 캡슐화를 이용한 항균 직물

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

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

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

박테리아 유래 항균 화합물의 효과를 평가하기 위한 지연 성장 억제 분석

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

신생 전염병 병원체 제거를위한 손씻기의 효능을 시험하는 방법

요약

더 많은 비디오 탐색

엔테로 바이러스 및 문화 RT-qPCR에 의해 물에서 노로 바이러스의 발생의 EPA 방법 1615 측정. 바이러스 샘플의 I. 컬렉션

온수 세탁의 바이러스 소독 효능 결정

잉크젯 프린터 기반으로 테스트 자동화 된 바둑판 배열 및 수동 시간 죽 일 방법 항균 시너지

Prion 안전 연구소 스와이프 테스트

도시 환경 면봉 - 환경 저수지에서 SARS-CoV-2의 샘플링 및 검출을 위한 파이프라인

정적 방법과 동적 방법을 결합한 접근법을 사용하여 낙농 생물막을 박멸하기 위한 유기 과산화산의 효능 평가

에센셜 오일의 나노 허브 캡슐화를 이용한 항균 직물

에센셜 오일의 나노 허브 캡슐화를 이용한 항균 직물

에센셜 오일의 나노 허브 캡슐화를 이용한 항균 직물

박테리아 유래 항균 화합물의 효과를 평가하기 위한 지연 성장 억제 분석