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中国防痨杂志 ›› 2024, Vol. 46 ›› Issue (5): 557-561.doi: 10.19982/j.issn.1000-6621.20240014

• 论著 • 上一篇    下一篇

应用低温等离子技术灭活体外结核分枝杆菌效果的初步研究

薛莲1, 剧猛1, 黄毅1, 赵国连2, 张语豪1, 王思翰1, 雷颖1, 党丽云3(), 左蕾1()   

  1. 1西安市胸科医院超声科,西安 710100
    2西安市胸科医院检验科,西安 710100
    3西安市胸科医院耐药结核科,西安 710100
  • 收稿日期:2024-01-10 出版日期:2024-05-10 发布日期:2024-04-29
  • 通信作者: 党丽云,Email:dangliyun@sina.com; 左蕾,Email:zuolei0221@163.com
  • 作者简介:注:剧猛与薛莲对本文有同等贡献,为并列第一作者
  • 基金资助:
    陕西省两链融合重点专项(2021LL-JB-06);陕西省自然科学基础研究计划(2022JQ-924);西安市科技计划项目(21YXYJ0001);西安市科技计划项目(2024JH-YLYB-0245);西安市卫生健康委员会科研项目(2023yb39)

A preliminary study of the in vitro bactericidal effect of low-temperature plasma technology on the Mycobacterium tuberculosis

Xue Lian1, Ju Meng1, Huang Yi1, Zhao Guolian2, Zhang Yuhao1, Wang Sihan1, Lei Ying1, Dang Liyun3(), Zuo Lei1()   

  1. 1Department of Ultrasound, Xi’an Chest Hospital, Xi’an 710100, China
    2Department of Laboratory Medicine, Xi’an Chest Hospital, Xi’an 710100, China
    3Department of Drug-resistant Tuberculosis, Xi’an Chest Hospital, Xi’an 710100, China
  • Received:2024-01-10 Online:2024-05-10 Published:2024-04-29
  • Contact: Dang Liyun, Email: dangliyun@sina.com; Zuo Lei, Email: zuolei0221@163.com
  • Supported by:
    Shaanxi Provincial Key Project of Two-Chain Integration(2021LL-JB-06);Shaanxi Provincial Natural Science Basic Research Program(2022JQ-924);Xi’an Science and Technology Plan Project(21YXYJ0001);Xi’an Science and Technology Plan Project(2024JH-YLYB-0245);Research Project of Xi’an Municipal Health Commission(2023yb39)

摘要:

目的: 初步探讨低温等离子技术在体外实验中对不同类型结核分枝杆菌的杀灭效果,旨在为结核分枝杆菌的杀灭提供新的技术和方法。方法: 将结核分枝杆菌标准株(H37Rv)、广泛耐药结核分枝杆菌(extensive drug-resistant Mycobacterium tuberculosis,XDR-MTB)、耐多药结核分枝杆菌(multidrug-resistant Mycobacterium tuberculosis,MDR-MTB)菌液接种在培养基上,于培养3周内配置成0.5麦氏浓度的菌悬液,再倍比稀释为试验用菌悬液,分别取2.0ml菌悬液样品置于无菌试管中;分别采用低温等离子系统PLACOAG(止血、凝固)模式对菌悬液行0、3、6、9、12、15s处理,随后放入培养箱,21d后分别计算各组不同作用时间存活的菌落数、杀菌对数值及杀菌率。结果: (1)H37Rv菌株(F=20.313,P=0.003)、XDR-MTB菌株(F=13.956,P=0.006)及MDR-MTB菌株(F=20.355,P=0.006)不同作用时间杀菌对数值差异均有统计学意义;3种菌株在作用9s时杀菌对数值比较差异有统计学意义(F=61.603,P=0.004),XDR-MTB菌株最低(0.50±0.00),H37Rv菌株居中(1.14±0.12),MDR-MTB菌株最高(1.23±0.00);3种菌株均于作用15s时杀菌对数值达到最大值,MDR-MTB菌株为1.71±0.00,H37Rv菌株为1.83±0.00,XDR-MTB菌株为1.35±0.21。(2)H37Rv菌株(F=10.458,P=0.012)、XDR-MTB菌株(F=10.945,P=0.011)及MDR-MTB菌株(F=9.424,P=0.015)不同作用时间杀菌率差异均有统计学意义;3种菌株在作用9s时杀菌率比较差异有统计学意义(F=287.890,P<0.001),XDR-MTB菌株最低[(68.25±0.00)%],H37Rv菌株居中[(92.59±2.10)%],MDR-MTB菌株最高[(94.12±0.00)%];3种菌株均于作用15s时杀菌率达到最大值,MDR-MTB菌株为(100.00±0.00)%,H37Rv菌株为(99.26±1.05)%,XDR-MTB菌株为(96.35±5.16)%。结论: 本研究通过体外实验证实了低温等离子技术对3种结核分枝杆菌均具有快速且明显的杀灭效果,但灭菌效果有所不同,需要制定个体化策略。

关键词: 分枝杆菌, 结核, 灭菌, 体外研究, 研究技术

Abstract:

Objective: Preliminary investigation of the killing effect of low-temperature plasma technology on different Mycobacterium tuberculosis in vitro experiments, aiming to provide a new technical method for killing common Mycobacterium tuberculosis. Methods: Mycobacterium tuberculosis H37Rv strain, extensive drug-resistant Mycobacterium tuberculosis (XDR-MTB) and multidrug-resistant Mycobacterium tuberculosis (MDR-MTB) mycobacterial fluids were inoculated on the culture medium, configured as a 0.5 mycobacterial suspensions within 3 weeks of incubation, and then multiply diluted to the test mycobacterial suspensions, and 2.0 ml of mycobacterial suspension samples were taken into sterile test tubes, the suspensions were ablated for 0, 3, 6, 9, 12 and 15 s by PLACOAG mode of low-temperature plasma surgery system, and then cultured in incubators. After 21 days, the number of the mean survival count, log10 reduction, and killing efficiency of each group at each treatment time point were calculated. Results: (1) The log10 reduction of H37Rv (F=20.313, P=0.003), XDR-MTB (F=13.956, P=0.006), and MDR-MTB (F=20.355, P=0.006) were statistically different at different treatment times. There was a statistically significant difference in the log10 reduction of three bacterial groups at 9 seconds (F=61.603, P=0.004), with XDR-MTB was the lowest (0.50±0.00), followed by H37Rv (1.14±0.12), and MDR-MTB was the highest (1.23±0.00), and the log10 reduction of the three groups reached the maximum value at 15 seconds, which were MDR-MTB (1.71±0.00), H37Rv (1.83±0.00), XDR-MTB (1.35±0.21). (2) The killing efficiency of H37Rv (F=10.458, P=0.012), XDR-MTB (F=10.945, P=0.011), and MDR-MTB (F=9.424, P=0.015) were statistically different at different treatment times. There was a statistically significant difference in the killing efficiency of three bacterial groups at 9 seconds (F=287.890, P<0.001), with XDR-MTB was the lowest ((68.25±0.00) %), followed by H37Rv ((92.59±2.10) %), and MDR-MTB was the highest ((94.12±0.00) %), and the killing efficiency of the three groups reached the maximum value at 15 seconds, which were MDR-MTB (100.00±0.00) %, H37Rv (96.26±1.05) %, XDR-MTB (99.35±5.16) %. Conclusion: In this study, in vitro experiments confirmed that low-temperature plasma technique has a rapid and significant killing effect on three Mycobacterium tuberculosis species, and that the killing effect varies and requires individualized strategy.

Key words: Mycobacterium tuberculosis, Sterilization, In vitro, Investigative techniques

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