变形链球菌耐氟菌株的筛选及分子鉴定

doi:10.13481/j.1671-587x.20140243

吉林大学学报(医学版)

变形链球菌耐氟菌株的筛选及分子鉴定

卢春英¹,张志民¹,于浩²，郝林琳²,张桐菲¹，李振玲^1,李朋莲¹,赵旭^1

1．吉林大学口腔医院牙体牙髓病科，吉林长春130021；2．吉林大学动物科学学院生物技术系，吉林长春130062

收稿日期:2013-10-28 出版日期:2014-03-28 发布日期:2014-05-28
通讯作者: 张志民（Tel：0431-85579500，E-mail：zhangzm1964@sina.com） E-mail:zhangzm1964@sina.com
作者简介:卢春英（1987-），女，山东省德州市人，在读医学硕士，主要从事龋病病因学基础及临床研究。
基金资助:
国家自然科学基金资助课题（3A411X463431）

Screening and molecular identification of fluoride-resistant strain of Streptococcus mutans

LU Chun-ying¹,ZHANG Zhi-min¹,YU Hao²,HAO Lin-lin²,ZHANG Tong-fei¹,LI Zhen-ling¹,LI Peng-lian¹,ZHAO Xu¹

1. Department of 　 Operative Dentistry and Endodontics,Stomatology Hospital,Jilin University,Changchun 130021,China;2. Department of Biotechnology,School of Animal Sciences,Jilin University, Changchun 130062,China

Received:2013-10-28 Online:2014-03-28 Published:2014-05-28

摘要/Abstract

摘要：

目的：探讨变形链球菌(S.mutans)获得耐氟性的培养条件及其鉴定方法，为鉴定变形链球菌耐氟菌株提供理论依据。方法：在微需氧条件(95%N2,5%CO2)下，以梯度氟化钠(NaF)浓度对变形链球菌UA159进行培养，获得耐氟菌株。从表现型和基因型上鉴定所培养菌株。观察菌株的表型特征；采用生理生化鉴定和PCR法对该菌株16S rDNA进行克隆鉴定。结果：微需氧（95%N2,5%CO2）条件下培养出耐氟变形链球菌株。形态学观察、生理生化鉴定和16S rDNA鉴定，该菌株具有耐氟稳定性，与变形链球菌UA159的16S rDNA序列同源性是100%,确定该菌株为变形链球菌耐氟菌株。结论：变形链球菌耐氟菌株培养条件是微需氧（95%N2,5%CO2）和脑心浸液(BHI)培养基中培养48 h。经NaF诱导的变形链球菌UA159鉴定为变形链球菌UA159的突变株变形链球菌UA159-FR。


关键词: 耐氟菌株, 分子鉴定, 龋病, 16S rDNA

Abstract:

Objective To probe the culture conditions and identification method of the fluoride-resistant strain of Streptococcus mutans(S.mutans)　and to provide theoretical basis for identifying fluoride-resistant strain of S.mutans.Methods In the microaerobic conditions(95%N2,5%CO2),the S.mutans UA159 were induced with NaF concentration gradient,and the fluoride-resistant strain of S.mutans was obtained.The bacteria was confirmed in terms of the phenotype and genotype.16S rDNA cloning identification of this bacteria was done by physiological and biochemical identification and PCR methods. The phenotypic characteristics of the strain was observed.Results In the microaerobic conditions(95%N2,5%CO2), the fluoride-resistant strain was obtained.The phenotypic characteristics observation,physiological and biochemical identification,and the identification of 16S rDNA results showed that this strain possessed fluoride-resistant stability,and the homology of this bacteria with S.mutans UA159 was 100%,and it was confirmed that this strain was the fluoride-resistant strain of S.mutans.Conclusion The culture conditions of fluoride-resistant strain of S.mutans are microaerobic conditions(95%N2,5%CO2) in the brain heart infusion(BHI) substrate for 48 h.S.mutans UA159 induced with NaF is identified as S.mutans UA159-FR.

Key words: , fluoride-resistant strain；molecular identification；tooth decay；16S rDNA

中图分类号:

卢春英,张志民,于浩，郝林琳,张桐菲，李振玲,李朋莲,赵旭. 变形链球菌耐氟菌株的筛选及分子鉴定[J]. 吉林大学学报(医学版), 2014, 40(02): 437-440.

LU Chun-ying,ZHANG Zhi-min,YU Hao,HAO Lin-lin,ZHANG Tong-fei,LI Zhen-ling, LI Peng-lian,ZHAO Xu. Screening and molecular identification of fluoride-resistant strain of Streptococcus mutans[J]. Journal of Jilin University Medicine Edition, 2014, 40(02): 437-440.

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变形链球菌耐氟菌株的筛选及分子鉴定

Screening and molecular identification of fluoride-resistant strain of Streptococcus mutans

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