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人诺如病毒替代物猫杯状病毒和鼠诺如病毒在地毯上的恢复优化及存活情况

Recovery Optimization and Survival of the Human Norovirus Surrogates Feline Calicivirus and Murine Norovirus on Carpet.

作者信息

Buckley David, Fraser Angela, Huang Guohui, Jiang Xiuping

机构信息

Department of Biological Sciences, Clemson University, Clemson, South Carolina, USA.

Department of Food, Nutrition, and Packaging Science, Clemson University, Clemson, South Carolina, USA.

出版信息

Appl Environ Microbiol. 2017 Oct 31;83(22). doi: 10.1128/AEM.01336-17. Print 2017 Nov 15.

Abstract

Carpets have been implicated in prolonged and reoccurring outbreaks of human noroviruses (HuNoV), the leading cause of acute gastroenteritis worldwide. Viral recovery from environmental surfaces, such as carpet, remains undeveloped. Our aim was to determine survival of HuNoV surrogates on an understudied environmental surface, carpet. First, we measured the zeta potential and absorption capacity of wool and nylon carpet fibers, we then developed a minispin column elution (MSC) method, and lastly we characterized the survival of HuNoV surrogates, feline calicivirus (FCV) and murine norovirus (MNV), over 60 days under 30 and 70% relative humidity (RH) on two types of carpet and one glass surface. Carpet surface charge was negative between relevant pH values (i.e., pH 7 to 9). In addition, wool could absorb approximately two times more liquid than nylon. The percent recovery efficiency obtained by the MSC method ranged from 4.34 to 20.89% and from 30.71 to 54.14% for FCV and MNV on carpet fibers, respectively, after desiccation. Overall, elution buffer type did not significantly affect recovery. Infectious FCV or MNV survived between <1 and 15 or between 3 and 15 days, respectively. However, MNV survived longer under some conditions and at significantly ( < 0.05) higher titers compared to FCV. Albeit, surrogates followed similar survival trends, i.e., both survived longest on wool then nylon and glass, while 30% RH provided a more hospitable environment compared to 70% RH. Reverse transcription-quantitative PCR signals for both surrogates were detectable for the entire study, but FCV genomic copies experienced significantly higher reductions (<3.80 log copies) on all surfaces compared to MNV (<1.10 log copies). Human noroviruses (HuNoV) are the leading cause of acute gastroenteritis worldwide. Classical symptoms of illness include vomiting and diarrhea which could lead to severe dehydration and death. HuNoV are transmitted by the fecal-oral or vomitus-oral route via person-to-person contact, food, water, and/or environmental surfaces. Published laboratory-controlled studies have documented the environmental stability of HuNoV on hard surfaces, but there is limited laboratory-based evidence available about survival on soft surfaces, e.g., carpet and upholstered furniture. Several epidemiological reports have suggested soft surfaces may be HuNoV fomites illustrating the importance of conducting a survival study. The three objectives of our research were to demonstrate techniques to characterize soft surfaces, develop a viral elution method for carpet, and characterize the survival of HuNoV surrogates on carpet. These results can be used to improve microbial risk assessments, the development of much-needed soft surface disinfectant, and standardizing protocols for future soft surface studies.

摘要

地毯与人类诺如病毒(HuNoV)的长期反复爆发有关,HuNoV是全球急性肠胃炎的主要病因。从地毯等环境表面恢复病毒的方法仍未得到充分发展。我们的目的是确定HuNoV替代病毒在研究较少的环境表面——地毯上的存活情况。首先,我们测量了羊毛和尼龙地毯纤维的zeta电位和吸附能力,然后开发了一种微型旋转柱洗脱(MSC)方法,最后我们对HuNoV替代病毒猫杯状病毒(FCV)和小鼠诺如病毒(MNV)在两种类型的地毯和一个玻璃表面上,于30%和70%相对湿度(RH)下60天内的存活情况进行了表征。在相关pH值(即pH 7至9)之间,地毯表面电荷为负。此外,羊毛比尼龙能多吸收约两倍的液体。干燥后,通过MSC方法获得的FCV和MNV在地毯纤维上的回收效率分别为4.34%至20.89%和30.71%至54.14%。总体而言,洗脱缓冲液类型对回收率没有显著影响。感染性FCV或MNV分别在<1至15天或3至15天内存活。然而,在某些条件下,MNV存活时间更长,且与FCV相比,其滴度显著(<0.05)更高。尽管替代病毒遵循相似的存活趋势,即两者在羊毛上存活时间最长,其次是尼龙和玻璃,而30%RH比70%RH提供了更适宜的环境。在整个研究中,两种替代病毒的逆转录定量PCR信号均可检测到,但与MNV(<1.10 log拷贝)相比,FCV基因组拷贝在所有表面上的减少幅度显著更高(<3.80 log拷贝)。人类诺如病毒(HuNoV)是全球急性肠胃炎的主要病因。疾病的典型症状包括呕吐和腹泻,这可能导致严重脱水和死亡。HuNoV通过粪口或呕吐物口途径,经人传人接触、食物、水和/或环境表面传播。已发表的实验室对照研究记录了HuNoV在硬表面上的环境稳定性,但关于其在软表面(如地毯和软垫家具)上存活的基于实验室的证据有限。几份流行病学报告表明,软表面可能是HuNoV的污染物,这说明了进行存活研究的重要性。我们研究的三个目标是展示表征软表面的技术、开发一种用于地毯的病毒洗脱方法,以及表征HuNoV替代病毒在地毯上的存活情况。这些结果可用于改进微生物风险评估、开发急需的软表面消毒剂,以及规范未来软表面研究的方案。

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