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傅玮涛,苏敏,吴初活,施炜,卢芳国,向琴,谭劲,周小青.福赛斯坦纳菌体外培养实验技术的研究[J].湖南中医药大学学报英文版,2018,38(1):43-46.[Click to copy
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福赛斯坦纳菌体外培养实验技术的研究 |
傅玮涛,苏敏,吴初活,施炜,卢芳国,向琴,谭劲,周小青 |
(湖南中医药大学医学院, 湖南 长沙 410208;湖南中医药大学中医诊断重点实验室, 湖南 长沙 410007;湖南中医药大学第一附属医院口腔科, 湖南 长沙 410007) |
摘要: |
目的 探索福赛斯坦纳菌生长繁殖的基本条件,比较N-乙酰胞壁酸及不同营养因子对其生长繁殖的影响,筛选该菌体外培养的最佳营养条件,建立其体外培养的技术方法。方法 (1)培养基制备:在血平板中加0.005% N-乙酰胞壁酸形成改良培养基;制备血平板与不同浓度(0.5、3、10、50、100 mg/L)的N-乙酰胞壁酸形成的培养基;制备10 mg/L N-乙酰胞壁酸与氯化血红素、酵母提取物、维生素K3、脱纤维羊血组合形成不同组别的培养基。(2)接种培养后,用磷酸盐缓冲液将细菌洗脱,并用酶标仪(OD600)测定其吸光度。结果 福赛斯坦纳菌在含N-乙酰胞壁酸的培养基中生长更好,浓度为10 mg/L时促进作用最明显且细菌形态更为典型。在组3培养基中生长最快。结论 N-乙酰胞壁酸能促进福赛斯坦纳菌的生长繁殖,浓度为10 mg/L时促进作用最明显;N-乙酰胞壁酸及氯化血红素、维生素K3、酵母提取物和脱纤维羊血组合能使其生长周期明显缩短。 |
关键词: 牙周炎 福赛斯坦纳菌 N-乙酰胞壁酸 体外培养 |
DOI:10.3969/j.issn.1674-070X.2017.01.011 |
Received:March 18, 2017 |
基金项目:2016年度湖南省大学生研究性学习和创新性实验计划项目(1021-001017094)。 |
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Investigation of Experimental Technique of Tannerella forsythia Cultured in Vitro |
FU Weitao,SU Min,WU Chuhuo,SHI Wei,LU Fangguo,XIANG Qin,TAN Jin,ZHOU Xiaoqing |
(Medical School, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Key Laboratory of TCM diagnosis, Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Stomatology Department, the First Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China) |
Abstract: |
Objective To investigate the basic requirements for the growth and proliferation of Tannerella forsythia and compare the growth-promoting effects with different nutritional factors and screen the optimum culture in vitro conditions and establish the technique of culturing it.Methods (1)Preparation of culture medium:0.005% N-acetylmuramic acid was added to the blood agar medium,forming the modified medium.Different concentration of N-acetylmuramic acid(0.5,3,10,50,100 mg/L)was added to the blood agar medium.The different mediums of N-acetylmuramic acid(10 mg/L)combined with other nutritional factors (chlorhematin,yeast extract,Vitamin K3,off fiber sheep blood) were formed.(2)After culturing,use PBS to elute the bacterial and measure its absorbance with enzyme-labelled instrument (OD600).Results Tannerella forsythia grew more better in the culture medium containing N-acetylmuramic acid,and the concentration at 10 mg/L was most obvious in growth-promoting effect and can lead to the change of bacterial morphology.The medium which consisted of hemin,yeast extract,menadione and sheep blood was the most suitable condition for its growth.Conclusion N-acetylmuramic acid can promote the growth of Tannerella forsythia,and the combination of N-acetylmuramic acid,hemin,menadione,yeast extract and sheep blood can shorten the growth cycle of it dramatically. |
Key words: periodontitis Tannerella forsythia N-acetylmuramic acid culture in vitro |
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