[1]谢娜,饶国洲,朱勇,等.单极射频低温等离子体杀菌的实验研究[J].现代检验医学杂志,2015,30(05):87-90.[doi:10.3969/j.issn.1671-7414.2015.05.026]
　XIE Na,RAO Guo-zhou,ZHU Yong,et al.Experimental Study on Monopole Radio Frequency Low Temperature Plasma Sterilization[J].Journal of Modern Laboratory Medicine,2015,30(05):87-90.[doi:10.3969/j.issn.1671-7414.2015.05.026]
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单极射频低温等离子体杀菌的实验研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第30卷
期数:
2015年05期
页码:
87-90
栏目:
论著
出版日期:
2015-12-10

文章信息/Info

Title:
Experimental Study on Monopole Radio Frequency Low Temperature Plasma Sterilization
作者:
谢娜1; 饶国洲2; 朱勇1; 李子夏1; 唐成芳1; 娄鸣1; 王丹杨1
1.西安医学院口腔医学系，西安710021；2.西安交通大学口腔医院，西安710004
Author(s):
XIE Na1; RAO Guo-zhou2; ZHU Yong1; LI Zi-xia1; TANG Cheng-fang1; LOU Ming1;  WANG Dan-yang1
1.Department of Stomatology，Xi’an Medical University，Xi’an 710021，China； 2.Hospital of Stomatology，Xi’an Jiaotong University，Xi’an 710004，China
关键词:
低温等离子体; 杀菌作用; 金黄色葡萄球菌; 大肠埃希菌; 超微结构
分类号:
R446.5
DOI:
10.3969/j.issn.1671-7414.2015.05.026
文献标志码:
A
摘要:
目的探讨单极射频低温等离子体在设定最佳参数下，作用不同时间的杀菌效果及机理。方法将金黄色葡萄球菌(ATCC25923)和大肠埃希菌(ATCC25922)标准菌株接种于营养琼脂平板，37℃进行活化培养24 h，挑取菌落以无菌生理盐水稀释成105 cfu/ml细胞悬液，取10 μl均匀涂布在无菌载玻片上，设对照组(照射前)和实验组(照射后)，每个作用时间设3个复片，采用单极射频低温等离子体装置(参数设置为10 W，10 KV，10 KHz，He/O2=2%，2 L/min)对其进行照射。照射时间分别为5，10，40，60，300，600，720和900s，通过菌落计数法计算杀灭率，检测不同照射时间的杀灭效果，电镜观察照射前后的细菌形态和超微结构的变化。结果对照组：金黄色葡萄球菌总数1 798 cfu/10 μl；大肠埃希菌总数2 563 cfu/10 μl。实验组：5，10，40，60，300，600，720和900s，其存活菌数和杀灭率分别为金黄色葡萄球菌：945 cfu/10 μl(47.4%)，823 cfu/10 μl(54.2%)，731 cfu/10 μl(59.3%)，586 cfu/10 μl(67.4%)，324 cfu/10 μl(81.9%)，107 cfu/10 μl(94.0%)，6 cfu/10 μl(99.7%)，0 cfu/10 μl(100%)；大肠埃希菌：1 546 cfu/10 μl(39.7%)，1 389 cfu/10 μl(45.8%)，1 282 cfu/10 μl(49.9%)，1 085 cfu/10 μl(57.7%)，579 cfu/10 μl(77.4%)，228 cfu/10 μl(91.7%)，11 cfu/10 μl(99.6%)和0 cfu/10 μl(100%)。结果表明不同照射时间其存活菌数和杀灭效果有所不同，其中照射900s时杀灭率达100%。电镜下观察可见金黄色葡萄球菌细胞壁破裂、模糊、溶解、胞浆内容物外流，失去完整性，胞内染色变浅，核质疏松；大肠埃希菌长度缩短、边缘粗糙、胞壁溶解、胞内出现空泡样改变。结论单极射频低温等离子体能有效地杀灭革兰氏阳性球菌和革兰氏阴性杆菌，其杀灭率随照射时间的延长而增加，其机理与电离产生的带电粒子破坏细菌胞壁、脂蛋白、脂多糖及DNA的作用有关。
Abstract:
ObjectiveTo investigate the effect and mechanism of action of monopole radio frequency low temperature plasma in the optimal parameters．MethodsStaphylococcus aureus (ATCC25923) and Escherichia coli (ATCC25922) strain was inoculated in standard nutrient agar plate，37℃ activated cultured 24 h．Colonies were picked with sterile normal saline and diluted to 105 cfu/ml cell suspension，taking 10 μl sterile evenly coated on a glass slide．The control group (before irradiation) and the experimental group (after irradiation)，each time setting the role of three complex pieces，were using low-temperature plasma unipolar radiofrequency device (parameter 10 W，10 KV，10 KHz，He/O2=2%，2 L/min) be irradiated．Irradiation time was 5，10，40，60，300，600，720，900 s and the sterilization rate was calculated by colony counting method to detect the effect of different exposure times．The morphology and ultrastructure of bacteria was detected by scanning electron microscope (SEM)．ResultsIn the control group：Staphylococcus aureus was total 1 798 cfu/10 μl and Escherichia coli was total 2 563 cfu/10 μl．In the experimental group：5，10，40，60，300，600，720，900s and the Staphylococcus aureus survival bacteria and sterilization rates was：945 cfu/10 μl (47.4%)，823 cfu/10 μl (54.2%)，731 cfu/10 μl (59.3%)，586 cfu/10 μl (67.4%)，324 cfu/10 μl (81.9%)，107 cfu/10 μl (94.0%)，6 cfu/10 μl (99.7%)，0 cfu/10μl (100%)．Escherichiacoli：1 546 cfu/10 μl (39.7%)，1 389 cfu/10 μl (45.8%)，1 282 cfu/10 μl (49.9%)，1 085 cfu/10 μl (57.7%)，579 cfu/10 μl (77.4%)，228 cfu/10 μl (91.7%)，11 cfu/10 μl (99.6%) and 0 cfu/10 μl (100%)．The results showed that different exposure times and sterilization bacteria survival effects were different，among 900s irradiation to kill up to 100%．Through SEM，observed the cytoderm of staphylococcus aureus turned to be distorted or ruptured．Similarly，the Escherichia coli cytoderm was decomposed partly，and vacuolar degeneration was also aggravated．ConclusionBoth Gram positive and negative bacteria could be sterilized by monopole radio frequency low temperature plasma．The sterilization rates were enhanced with the extended exposure time．The mechanism may be related with the charged particle produced by the ionization of plasma，which causes damage to cytoderm，lipoprotein，lipopolysaccharide and DNA．

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备注/Memo

备注/Memo:
基金项目：陕西省卫生厅科研项目(2014D19)，陕西省教育厅2012年科学研究项目(12JK0757)，陕西省教育厅专项科研计划项目(2013JK0784)。 作者简介：谢娜(1983-)，女，硕士，口腔系助教，主要从事教学及干细胞研究，E-mail：springna888@163.com。 通讯作者：饶国洲，男，主任检验师(教授)，硕士生导师，主要从事分子与细胞生物学研究。
更新日期/Last Update: 1900-01-01