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中华临床医师杂志(电子版) ›› 2024, Vol. 18 ›› Issue (11) : 1037 -1043. doi: 10.3877/cma.j.issn.1674-0785.2024.11.010

基础研究

低温等离子消融技术对体外常见病菌的杀菌效果研究
常哲1, 党丽云2, 黄毅3, 左蕾3, 薛莲3, 赵国连4, 任斐2, 牛俣鑫4, 高良5,()   
  1. 1.710000 西安,西安市凤城医院超声科
    2.710100 西安,西安市胸科医院耐药结核科
    3.710100 西安,西安市胸科医院超声科
    4.710100 西安,西安市胸科医院检验科
    5.710004 西安,西安市人民医院(西安市第四医院)超声科
  • 收稿日期:2024-10-21 出版日期:2024-11-15
  • 通信作者: 高良
  • 基金资助:
    陕西省两链融合重点专项(No.2021LL-JB-06)西安市卫生健康委员会科研项目(No.2023yb39)西安英才计划项目(No.XAYC220043)

In vitro bactericidal effect of low-temperature plasma ablation on common pathogens

Zhe Chang1, Liyun Dang2, Yi Huang3, Lei Zuo3, Lian Xue3, Guolian Zhao4, Fei Ren2, Yuxin Niu4, Liang Gao5,()   

  1. 1.Department of Ultrasound, Xi'an Fengcheng Hospital, Xi’an 710000, China
    2.Department of Drug-resistant Tuberculosis, Xi’an Chest Hospital, Xi’an 710100,China
    3.Department of Ultrasound, Xi’an Chest Hospital, Xi’an 710100,China
    4.Department of Laboratory Medicine, Xi’an Chest Hospital, Xi’an 710100,China
    5.Department of Ultrasound, Xi'an People’s Hospital (Xi’an Fourth Hospital), Xi'an 710004, China
  • Received:2024-10-21 Published:2024-11-15
  • Corresponding author: Liang Gao
引用本文:

常哲, 党丽云, 黄毅, 左蕾, 薛莲, 赵国连, 任斐, 牛俣鑫, 高良. 低温等离子消融技术对体外常见病菌的杀菌效果研究[J/OL]. 中华临床医师杂志(电子版), 2024, 18(11): 1037-1043.

Zhe Chang, Liyun Dang, Yi Huang, Lei Zuo, Lian Xue, Guolian Zhao, Fei Ren, Yuxin Niu, Liang Gao. In vitro bactericidal effect of low-temperature plasma ablation on common pathogens[J/OL]. Chinese Journal of Clinicians(Electronic Edition), 2024, 18(11): 1037-1043.

目的

初步探讨低温等离子消融技术在体外实验中对不同病菌的杀灭效果,旨在为常见病菌的杀灭提供新的技术方法。

方法

将金黄色葡萄球菌、大肠埃希菌、铜绿假单胞菌以及白色念珠菌菌液接种在培养基上,培养至对数期,配置为0.5 麦氏菌悬液,分别取2.0 ml 菌悬液样品置于无菌试管中,采用低温等离子消融手术系统的PLACOAG 模式对菌悬液分别进行0 s、3 s、6 s、9 s、12 s、15 s、18 s 的消融处理,随后放入培养箱培养,并分别计算各组各消融时间点存活的菌落数、杀菌对数值及杀菌率。

结果

金黄色葡萄球菌(F=40.360,P<0.001)、大肠埃希菌(F=72.568,P<0.001)、铜绿假单胞菌(F=129.028,P<0.001)以及白色念珠菌(F=85.744,P<0.001)不同处理时间菌落计数差异均具有统计学意义;4 个病菌组菌落计数均在消融18s 达到最小值,4 组比较差异具有统计学意义(F=27.105,P<0.001),其中白色念珠菌最少,大肠埃希菌和铜绿假单胞菌次之,金黄色葡萄球菌最多;金黄色葡萄球菌(F=56.287,P<0.001)、大肠埃希菌(F=36.529,P<0.001)、铜绿假单胞菌(F=22.853,P<0.001)以及白色念珠菌(F=133.974,P<0.001)不同处理时间平均杀灭对数值差异均具有统计学意义;4 个病菌组平均杀灭对数值均在消融18 s 达到最大值,4 组比较差异具有统计学意义(F=171.764,P<0.001),其中白色念珠菌最大,大肠埃希菌和铜绿假单胞菌次之,金黄色葡萄球菌最小;金黄色葡萄球菌(F=86.522,P<0.001)、大肠埃希菌(F=46.388,P<0.001)、铜绿假单胞菌(F=62.503,P<0.001)以及白色念珠菌(F=51.726,P<0.001)不同处理时间病菌致死率差异均具有统计学意义;金黄色葡萄球菌、大肠埃希菌和铜绿假单胞菌病菌致死率在消融18s 达到最大值,白色念珠菌在12 s 即达到最大值,4 个病菌组在消融18 s 病菌致死率差异具有统计学意义(F=20.607,P<0.001);其中白色念珠菌和铜绿假单胞菌病最高,大肠埃希菌次之,金黄色葡萄球菌最低。

结论

本研究通过体外实验证实了低温等离子消融技术对常见的病菌均具有快速且明显的杀灭效果。四种常见病菌灭菌效果有所不同,需要制定个体化的消融策略。

Objective

To preliminarily explore the killing effect of low-temperature plasma ablation on different pathogens in vitro, aiming to provide a new technical method for killing common pathogens.

Methods

Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Candida albicans were inoculated into culture media, cultured to the logarithmic growth stage, and prepared as 0.5 McFarland bacterial suspensions, respectively. Bacterial suspension samples (2.0 mL) were placed in sterile test tubes, and the PLACOAG mode of the low-temperature plasma ablation system was used for ablation treatments on the bacterial suspensions for 0 s, 3 s, 6 s, 9 s, 12 s, 15 s, and 18 s, respectively. Then, the samples were placed in an incubator for cultivation.The mean survival count, bactericidal logarithms, and bactericidal rate at each ablation time point in each group were calculated.

Results

The colony counts of Staphylococcus aureus (F=40.360, P<0.001), Escherichia coli (F=72.568, P<0.001), Pseudomonas aeruginosa (F=129.028, P<0.001), and Candida albicans (F=85.744, P<0.001) were statistically different among different treatment durations. The colony counts of the four pathogen groups all reached the minimum at 18 s of ablation, but they were statistically different among the four groups (F=27.105, P<0.001), with that of Candida albicans being the least, those of Escherichia coli and Pseudomonas aeruginosa being the second,and that of Staphylococcus aureus being the most. The average killing logarithm values for Staphylococcus aureus (F=56.287, P<0.001), Escherichia coli (F=36.529, P<0.001), Pseudomonas aeruginosa (F=22.853,P<0.001), and Candida albicans (F=133.974, P<0.001) were statistically different among different treatment durations. The average killing logarithm values for the four pathogen groups reached the maximum value at 18 s of ablation, but they were statistically different among the four groups (F=171.764, P<0.001), with that for Candida albicans being the largest, those for Escherichia coli and Pseudomonas aeruginosa being the second largest, and that for Staphylococcus aureus being the smallest. The average maximum bactericidal rate for Staphylococcus aureus (F=86.522, P<0.001), Escherichia coli (F=46.388, P<0.001), Pseudomonas aeruginosa (F=62.503, P<0.001), and Candida albicans (F=51.726, P<0.001) were statistically different among different treatment durations. The maximum bactericidal rates for Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa reached their maximum values at 18 s of ablation, while that for Candida albicans reached its maximum at 12 s. There was a statistical difference in the bactericidal rates among the four treatment groups at 18s of ablation (F=20.607, P<0.001), with those for Candida albicans and Pseudomonas aeruginosa being the highest, followed by that for Escherichia coli, and that for Staphylococcus aureus being the lowest.

Conclusion

This study confirmed that low-temperature plasma ablation has a rapid and significant killing effect on common pathogens in vitro. The killing effect on the four common pathogens is variable and individualisation of the ablation strategy is thus required.

图1 低温等离子消融设备及方法。图a 为低温等离子体手术系统PLA-600;图b 为MC247 等离子刀头(直径0.9 mm/长度20 cm);图c 为2.0 ml 菌悬液样品置于无菌试管中,调节等离子刀头置于试管底部上方约10 mm 处进行消融,处理时间分别为0 s、3 s、6 s、9 s、12 s、15 s、18 s
表1 四种常见病菌行低温等离子消融不同处理时间组的菌落计数比较(±s
图2 低温等离子消融四种病菌的体外杀菌效果
表2 四种常见病菌行低温等离子消融不同处理时间的平均杀灭对数值比较
表3 四种常见病菌行低温等离子消融不同处理时间的病菌致死率(%)比较
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