口腔综合治疗台水路系统中假单胞菌属污染状况的检测

发布时间：2018-02-20 01:46

本文关键词： 口腔综合治疗台水路系统假单胞菌属 Miseq测序技术　出处：《南京医科大学学报(自然科学版)》2017年04期 　论文类型：期刊论文

【摘要】：目的:采用细菌培养技术与Miseq测序技术检测牙科综合治疗台水路系统(dental unit waterlines,DUWLs)中的假单胞菌属,以综合评估假单胞菌属在DUWLs中的污染情况。方法:选择南京医科大学附属口腔医院25台口腔综合治疗台作为研究对象。在医院开诊前收集三用枪水样400 mL,平均分为2份,一份用于细菌培养。另一份用于提取细菌基因组总DNA,经细菌通用引物PCR扩增后构建宏基因组文库,用于Miseq测序和生物信息学分析。结果:25个样本在不同浓度条件下经细菌培养法均未检出假单胞菌属。16个样本成功构建宏基因组文库,经Miseq测序均检测到假单胞菌属存在,检出率为100%,假单胞菌属在16个样本中的丰度为(3.02±2.12)%。结论:DUWLs中存在低水平的假单胞菌属污染,但假单胞菌属大部分可能处于死亡或"活的非可培养"状态,不具有较大的生物危害性。与传统细菌培养法相比,Miseq测序技术在特定病原菌的检测方面表现出了较高的敏感性,但无法区分样本中细菌的活性,若要将高通量测序技术用于活性病原菌的检测还需要进一步的研究。
[Abstract]:Objective: to detect Pseudomonas in dental unit waterlines by bacterial culture and Miseq sequencing. The contamination of Pseudomonas in DUWLs was evaluated synthetically. Methods: 25 oral comprehensive treatment sets were selected from the Stomatology Hospital affiliated to Nanjing Medical University as the research objects. The water samples of three guns were collected before the opening of the hospital, which were divided into 2 parts on average. One was used for bacterial culture, the other was used to extract the total genomic DNA of bacteria, and the macro genome library was constructed by amplification of bacterial universal primer PCR. Miseq sequencing and bioinformatics analysis. Results: Pseudomonas was not detected in 25 samples under different concentration conditions. Sixteen samples were successfully constructed macrogenomic library, and Pseudomonas were detected by Miseq sequencing. The detection rate was 100 and the abundance of Pseudomonas in 16 samples was 3.02 卤2.12.Conclusion there is a low level of Pseudomonas contamination in the 20% DUWLs, but most of the Pseudomonas may be dead or "live non-culturable". Compared with the traditional bacterial culture method, the Miseq sequencing technique showed a high sensitivity in the detection of specific pathogens, but could not distinguish the activity of bacteria in the samples. If high-throughput sequencing is to be used for the detection of active pathogens, further research is needed.
【作者单位】：南京医科大学口腔疾病研究江苏省重点实验室南京医科大学附属口腔医院牙体牙髓科;
【基金】：国家自然科学基金(81300868) 江苏省高校大学生实践创新训练计划(201410312010Z)
【分类号】：R378.991

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