本文關(guān)鍵詞： 口腔綜合治療臺(tái)水路系統(tǒng) 假單胞菌屬 Miseq測序技術(shù)　出處：《南京醫(yī)科大學(xué)學(xué)報(bào)(自然科學(xué)版)》2017年04期 　論文類型：期刊論文

【摘要】：目的:采用細(xì)菌培養(yǎng)技術(shù)與Miseq測序技術(shù)檢測牙科綜合治療臺(tái)水路系統(tǒng)(dental unit waterlines,DUWLs)中的假單胞菌屬,以綜合評估假單胞菌屬在DUWLs中的污染情況。方法:選擇南京醫(yī)科大學(xué)附屬口腔醫(yī)院25臺(tái)口腔綜合治療臺(tái)作為研究對象。在醫(yī)院開診前收集三用槍水樣400 mL,平均分為2份,一份用于細(xì)菌培養(yǎng)。另一份用于提取細(xì)菌基因組總DNA,經(jīng)細(xì)菌通用引物PCR擴(kuò)增后構(gòu)建宏基因組文庫,用于Miseq測序和生物信息學(xué)分析。結(jié)果:25個(gè)樣本在不同濃度條件下經(jīng)細(xì)菌培養(yǎng)法均未檢出假單胞菌屬。16個(gè)樣本成功構(gòu)建宏基因組文庫,經(jīng)Miseq測序均檢測到假單胞菌屬存在,檢出率為100%,假單胞菌屬在16個(gè)樣本中的豐度為(3.02±2.12)%。結(jié)論:DUWLs中存在低水平的假單胞菌屬污染,但假單胞菌屬大部分可能處于死亡或"活的非可培養(yǎng)"狀態(tài),不具有較大的生物危害性。與傳統(tǒng)細(xì)菌培養(yǎng)法相比,Miseq測序技術(shù)在特定病原菌的檢測方面表現(xiàn)出了較高的敏感性,但無法區(qū)分樣本中細(xì)菌的活性,若要將高通量測序技術(shù)用于活性病原菌的檢測還需要進(jìn)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 鹵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.
【作者單位】： 南京醫(yī)科大學(xué)口腔疾病研究江蘇省重點(diǎn)實(shí)驗(yàn)室南京醫(yī)科大學(xué)附屬口腔醫(yī)院牙體牙髓科;
【基金】：國家自然科學(xué)基金(81300868) 江蘇省高校大學(xué)生實(shí)踐創(chuàng)新訓(xùn)練計(jì)劃(201410312010Z)
【分類號】：R378.991

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