Wolfe Marlene K, Lantagne Daniele S
Department of Civil and Environmental Engineering, Tufts University;
Department of Civil and Environmental Engineering, Tufts University.
J Vis Exp. 2017 Jun 7(124):55604. doi: 10.3791/55604.
Handwashing is widely recommended to prevent infectious disease transmission. However, little comparable evidence exists on the efficacy of handwashing methods in general. Additionally, little evidence exists comparing handwashing methods to determine which are most efficacious at removing infectious pathogens. Research is needed to provide evidence for the different approaches to handwashing that may be employed during infectious disease outbreaks. Here, a laboratory method to assess the efficacy of handwashing methods at removing microorganisms from hands and their persistence in rinse water is described. Volunteers' hands are first spiked with the test organism and then washed with each handwashing method of interest. Generally, surrogate microorganisms are used to protect human subjects from disease. The number of organisms remaining on volunteers' hands after washing is tested using a modified "glove juice" method: the hands are placed in gloves with an eluent and are scrubbed to suspend the microorganisms and make them available for analysis by membrane filtration (bacteria) or plaque assay (viruses/bacteriophages). Rinse water produced from the handwashing is directly collected for analysis. Handwashing efficacy is quantified by comparing the log reduction value between samples taken after handwashing to samples with no handwashing. Rinse water persistence is quantified by comparing rinse water samples from various handwashing methods to samples collected after handwashing with just water. While this method is limited by the need to use surrogate organisms to preserve the safety of human volunteers, it captures aspects of handwashing that are difficult to replicate in an in vitro study and fills research gaps on handwashing efficacy and the persistence of infectious organisms in rinse water.
广泛推荐洗手以预防传染病传播。然而,总体而言,关于洗手方法效果的可比证据很少。此外,比较不同洗手方法以确定哪种方法在去除传染性病原体方面最有效的证据也很少。需要开展研究,为传染病暴发期间可能采用的不同洗手方法提供证据。在此,描述了一种实验室方法,用于评估洗手方法在去除手上微生物及其在冲洗水中的持久性方面的效果。首先在志愿者手上接种测试微生物,然后用每种感兴趣的洗手方法进行洗手。一般来说,使用替代微生物来保护人类受试者免受疾病侵害。洗手后,使用改良的“手套汁液”方法检测志愿者手上残留的微生物数量:将手放入装有洗脱液的手套中,擦拭以悬浮微生物,使其可通过膜过滤(细菌)或噬斑测定(病毒/噬菌体)进行分析。直接收集洗手产生的冲洗水进行分析。通过比较洗手后采集的样本与未洗手样本之间的对数减少值来量化洗手效果。通过将各种洗手方法的冲洗水样本与仅用水洗手后采集的样本进行比较来量化冲洗水的持久性。虽然这种方法受到需要使用替代生物来保障人类志愿者安全的限制,但它捕捉到了在体外研究中难以复制的洗手方面的情况,并填补了关于洗手效果和冲洗水中传染性生物持久性的研究空白。