【作者】 马玉莹； 芦昕； 张利娟； 刘育含； 李帆； 谭凯璇； 张颖； 李修珍； 杨芳；

【Author】 Yuying Ma;Xin Lu;Lijuan Zhang;Yuhan Liu;Fan Li;Kaixuan Tan;Ying Zhang;Xiuzhen Li;Fang Yang;Stomatology Center, Qingdao Municipal Hospital;School of Stomatology, Dalian Medical University;Stomatology Department, Women & Children’s Health Care Hospital of Linyi;School of Stomatology, Qingdao University;

【机构】 青岛市市立医院口腔医学中心； 大连医科大学口腔医学院； 临沂市妇幼保健院口腔科； 青岛大学口腔医学院；

【摘要】 目的:系统评价重水拉曼技术在耐药性研究领域的可行性及次氯酸钠(Sodium hypochlorite,NaOCl)对粪肠球菌(Enterococcus faecalis,E.faecalis)的抑菌效能。方法:1)采用吸光度测定、重水拉曼技术评价粪肠球菌在不同浓度重水中生长状态及对重水的吸收规律,评价重水拉曼技术在细菌耐药性研究方面的普适性。2)采用肉汤稀释法,结合吸光度测定次氯酸钠对粪肠球菌的最小抑菌浓度(Minimum Inhibitory Concentration,MIC);采用重水拉曼技术(D2O-Labeled Single-Cell Raman Micro-spectroscopy)测定其对粪肠球菌的最低抑制代谢浓度(Minimum Inhibitory Concentration based on Metabolic Activity,MIC-MA)。结果:1)重水浓度≤40%时,粪肠球菌的正常生长不会受到抑制(P>0.05);粪肠球菌能够活跃代谢重水并在拉曼图谱特定区域出现重水峰,且重水峰面积与所加入重水浓度成线性正相关关系(R2=0.9585,P<0.01)。2)次氯酸钠对粪肠球菌的MIC值为:0.45g/L;MIC-MA值为:0.9g/L。次氯酸钠作用于粪肠球菌的MIC-MA为MIC的2倍。结论:重水拉曼技术在抗菌剂筛选及抗菌剂作用效能评价方面具有重要价值,适用于评价药物对细菌代谢活性的影响。次氯酸钠对粪肠球菌表现出明显的抑制作用。粪肠球菌细胞在MIC浓度作用下生长虽受到抑制,但仍存在代谢活性;而绝大部分粪肠球菌细胞的代谢活性在MIC-MA浓度作用下才得到抑制。更多还原

【Abstract】 Objectives:Systematic evaluation the feasibility of D₂O-labeled single-cell Raman micro-spectroscopy in drug resistance research and test the susceptibility of Sodium hypochlorite against Efaecalis.Methods:1) The growth of Efaecalis in different doses of D₂O and the regularity of D₂O intake were evaluated by absorbance measurement and D₂O-labeled single-cell Raman micro-spectroscopy,to evaluate the universality of D₂O-labeled single-cell Raman micro-spectroscopy in the study of bacterial resistance.2)Broth dilution method and absorbance measurement was performed to determined the Minimum Inhibitory Concentration（MIC） of NaClO for E.faecalis,and Minimum Inhibitory Concentration based on Metabolic Activity（MIC-MA） in vitro based on D₂O-Labeled Single-Cell Raman Micro-spectroscopy.Results:1) The concentration of D₂O in the medium at concentrations ≤40% did not have significant inhibition on the growth of E.faecalis,and E.faecalis can actively metabolize D₂O and appear C-D ratio in specific areas of Raman micro-spectroscopy results.Moreover,the C-D ratio of E.faecalis at the stationary phase was positively correlated with D₂O concentration.2)The MIC of NaClO for E.faecalis were 0.45 g/L;the MIC-MA of it were 0.9 g/L,and the concentration of MIC-MA is twice that of MIC.Conclusion:D₂O-labeled single-cell Raman micro-spectroscopy is of great value in screening antimicrobial agents and evaluating the efficacy of antimicrobial agents,which is suitable for evaluating the effect of drugs on bacterial metabolic activity.NaClO showed an effective antimicrobial activity against Efaecalis.E.faecalis ceased propagation yet remained metabolically highly active when it was exposed to the doses of NaClO at MIC level.Moreover,the metabolic activity of most cells was inhibited only when exposure to NaClO at MIC-MA level.更多还原