基于叠氮溴化丙锭结合高通量测序技术分析口腔综合治疗台水路系统中活菌多样性的研究
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

南京医科大学2017校基金(2017NJMU112);南京医科大学校基金(NMUB2019073)


Analysis on viable bacterial diversity in dental unit waterlines based on propidium monoazid in conjunction with high⁃throughput sequencing
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    目的:利用叠氮溴化丙锭(propidium monoazid,PMA)预处理方法结合高通量测序技术检测口腔综合治疗台水路系统(dental unit waterline,DUWL)中活菌微生物群落,以综合评估不同科室DUWL中的污染情况及探究活菌群落构成的多样性。方法:选择南京医科大学附属口腔医院33台口腔综合治疗台,牙体牙髓病科23台(E组),牙周病科10台(P组),在医院开诊前收集三用枪水样,PMA预处理水样后提取活菌总DNA,经细菌通用引物PCR扩增后构建宏基因组文库,用于高通量测序和生物信息学分析。结果:样本使用1 μL PMA(最终浓度20 μmol/L)预处理10 min后暴露于卤素光照射5 min,最适合检测DUWL样品中活菌的量;高通量测序显示DUWL中活菌微生物群落呈多样性,并具有潜在的细菌致病序列。科室间具有统计学差异的菌门为蓝藻菌门(Cyanobacteria)、放线菌门(Actinobacteria)、衣原体门(Chlamydiae)、Aerophobetes、绿弯菌门(Chloroflexi)、Bacteria_d_norank_k__unclassified、TM6(P < 0.05);属水平上,军团菌属(Legionella)、Methyloversatilis、Obscuribacterales__ norank差异有统计学意义(P < 0.05)。结论:DUWL中致病微生物的存在对牙科医务工作者和患者都具有潜在感染风险,应重视DUWL细菌群落的多样性,有必要对DUWL进行定期水质控制。

    Abstract:

    Objective:In order to evaluate the microbial contamination of different departments and explore the diversity of living bacterial community,the method of propidium monoazid(PMA) pretreatment combined with high-throughput sequencing technology was used to detect the living microbial community in the dental unit waterline(DUWL). Methods:A total of 33 dental chair units(DCU)were selected from the Affiliated Stomatological Hospital of Nanjing Medical University,with 23 from endodontics department(group E)and 10 from periodontics department(group P). Before patients arrived,water samples were collected from each DCU using a three-in-one air/water syringe. After PMA pretreatment,the total DNA of living bacteria was extracted. After PCR amplification with bacterial universal primers,the macrogenomic library was constructed for high-throughput sequencing and bioinformatics analysis. Results:Samples were pretreated with 1 μL of PMA(20 μmol/L)for 10 min and subsequently exposed to light irradiation for 5 min,which were optimal for detecting viable bacterial in samples of DUWL. The output water of DUWL was associatied with a high bacterial diversity and identified potential bacterial pathogenic sequence. The statistically different phyla between the two groups were Cyanobacteria,Actinobacteria,Chlamydiae,Aerophobetes,Chloroflexi,Bacteria_d_norank_k__unclassified,TM6(P < 0.05). The statistically different genera between the two groups were Legionella,Methyloversatilis,Obscuribacterales__norank(P < 0.05). Conclusion:The occurrence of potentially pathogenic microorganisms in these DUWLs demonstrated a potential infectious risk for both dental health care personnel and patients. Attention should be paid to the diversity of DUWL bacterial communities . It is necessary to proceed to regular water quality control of DUWLs.

    参考文献
    相似文献
    引证文献
引用本文

周如玉,平逸帆,张 元,王 娟.基于叠氮溴化丙锭结合高通量测序技术分析口腔综合治疗台水路系统中活菌多样性的研究[J].南京医科大学学报(自然科学版),2021,(6):847-855

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2020-04-01
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2021-06-26
  • 出版日期:
通知关闭
郑重声明