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洗手产品除病毒效果离体实验方法研究

In vitro method for evaluation on virus-eliminating effectiveness of hand wash agents: an experimental study
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摘要 目的 建立一种安全、简便和费用低廉的测试洗手及洗手产品除病毒效果的离体试验方法。方法 以噬菌体为替代病毒,仿真皮为载体设计试验流程,分析影响试验结果的要素,通过数据分析,研究洗手产品回收稀释液种类、噬菌体初始接种浓度及回收方式对结果稳定性的影响。使用不同洗手产品对多种噬菌体分别进行除病毒效果测试后进行统计分析,确定方法的适用性。结果 以噬菌体 φX174为污染病毒,0.1%蛋白胨生理盐水做为香皂和洗手液的回收稀释液使用时,其噬菌体的回收值为5.57和5.54,明显高于改良李氏肉汤吐温培养基(MLBT)、多用中和剂(PVUN)及磷酸缓冲液(PBS);对手摇瓶子2 min、摇床200 r/min振摇2 min、均质器标准强度快速拍打2 min后测得的噬菌体回收量进行单因素方差分析,P0.05=0.72,表明3者无统计学差异,但是在使用均质器拍打时的变异系数(CV)为0.74%最小,数据最稳定;4个初始接种浓度中,只有在107 pfu/mL与108 pfu/mL的浓度下,平皿上的噬菌体数量符合实验设计,超过4 Log10,对这2个浓度实验结果进行t检验,其P0.05=0.003,结合回收数据,认为108 pfu/mL是最合适的加样浓度;使用上述参数,分别以噬菌体MS2、φX174、φ6为污染病毒,使用香皂、洗手液及清水进行洗手除病毒评价,所有结果CV均<15%。结论 采用108 pfu/mL的噬菌体悬液污染仿真皮后使用洗手产品模拟洗手,均质器拍打2 min、0.1%蛋白胨生理盐水进行回收稀释,实验结果最稳定。多次重复实验结果显示,该方法适用于多种噬菌体,也适用于多种洗手产品,是一种简单可行的洗手及洗手产品除病毒效果评价方法,具有较高的应用价值。 Objective To develop a simple, safe, and low cost in vitro method for the evaluation on virus-eliminating effectiveness of hand-washing and hand wash agents.Methods We used artificial skin as human skin surrogate and bacteriophage as virus surrogate to construct an in vitro model. We analyzed effects of various recovery diluent, initial concentration of bacteriophage inoculation, and recovery method on detection reproducibility of the method to be established. Three bacteriophage strains were used in evaluations of bar and liquid soap. Statistical analyses on experimental data were performed to assess the applicability of the method.Results The recovery (logarithmic value) of φX174 bacteriophage was 5.57 and 5.54 for washing procedure with bar soap and liquid soap when using physiological saline with 0.1% peptone as the diluent and the values were obviously higher than those when using modified Letheen broth plus Tween (MLBT), polyvalent universal neutralizer (PVUN) or phosphate buffer saline (PBS) as the diluent. The results of one-way analysis of variance demonstrated that there was no significant difference in the recovery of bacteriophage among the three patterns of simulated 2-minute hand-washing (shaking the bottle manually, shaking violently with a shaker at 200 revolutions per minute or with a homogenizer) (P0.05=0.72);however, the variation in the recovery of bacteriophage was the lowest (coefficient of variation[CV]=0.74%) when using homogenizer in the simulated test. The bacteriophage plate count consistent with the experimental design (baseline recovery > 4 Log10, Pstudent-t test=0.003) was achieved under two (108 and 107 plaque forming unit[pfu]/ml) of the four initial concentrations of bacteriophage inoculation;while, the simulated virus-eliminating rate was significantly higher with the initial inoculation concentration of 108 pfu/ml than with that of 107 pfu/ml, indicating that 108 pfu/ml was the most optimal initial inoculation concentration. All the CV of variation in bacteriophage recovery wer
作者 孙廷丽 周少璐 杨秀茳 文霞 黎玉莲 谢小保 SUN Ting-li;ZHOU Shao-lu;YANG Xiu-jiang(Guangdong Institute of Microbiology, Guangdong Detection Center of Microbiology, State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory for Microbiology Culture Collection and Application, Guangdong Open Laboratory of Applied Microbiology, Guangzhou, Guangdong Province 510070, China)
出处 《中国公共卫生》 CAS CSCD 北大核心 2019年第1期42-47,共6页 Chinese Journal of Public Health
关键词 离体洗手模型 洗手 洗手产品 替代病毒 除病毒效果 噬菌体 in vitro hand-washing model hand wash hand wash agents surrogate virus virus-eliminating effectiveness bacteriophage
作者简介 孙廷丽(1982-),女,山东惠民人,助理研究员,硕士,研究方向:工业材料及产品(日化产品)抗生物性能检测方法及标准化研究.
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