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北京大学学报（医学版） ›› 2026, Vol. 58 ›› Issue (1): 50-59. doi: 10.19723/j.issn.1671-167X.2026.01.007

• 论著 • 上一篇    下一篇

1例成年猛性龋患者龈上菌斑中小韦荣球菌的分离及代谢特性

何梓玉1, 张辉1, 陈智滨2, 邢海霞1,*(), 潘洁1,*()   

  1. 1. 北京大学口腔医学院·口腔医院综合治疗科, 国家口腔医学中心, 国家口腔疾病临床医学研究中心, 口腔生物材料和数字诊疗装备国家工程研究中心, 北京 100081
    2. 北京大学口腔医学院·口腔医院牙周科, 国家口腔医学中心, 国家口腔疾病临床医学研究中心, 口腔生物材料和数字诊疗装备国家工程研究中心, 北京 100081
  • 收稿日期:2025-10-10 出版日期:2026-02-18 发布日期:2025-12-02
  • 通讯作者: 邢海霞, 潘洁
  • 基金资助:
    国家自然科学基金(51801003)

Isolation, identification, and metabolic characterization of a Veillonella parvula isolated from supragingival plaque in a patient with rampant caries

Ziyu HE1, Hui ZHANG1, Zhibin CHEN2, Haixia XING1,*(), Jie PAN1,*()   

  1. 1. Department of General Dentistry, Peking University School and Hospital of Stomatology & National Center for Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices, Beijing 100081, China
    2. Department of Periodontology, Peking University School and Hospital of Stomatology & National Center for Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices, Beijing 100081, China
  • Received:2025-10-10 Online:2026-02-18 Published:2025-12-02
  • Contact: Haixia XING, Jie PAN
  • Supported by:
    the National Natural Science Foundation of China(51801003)

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摘要：

目的: 从成年猛性龋患者健康牙面的龈上菌斑中分离并培养野生株小韦荣球菌(Veillonella parvula, V. parvula), 研究其生长及代谢特性, 初步探究其与变异链球菌(Streptococcus mutans, S. mutans)的相互作用。方法: 利用韦荣氏球菌琼脂培养基, 从成年猛性龋患者健康牙面的龈上菌斑中分离并纯化野生株V. parvula, 进行形态、生化和16S rRNA基因鉴定。取标准株V. parvula作为对照组, 用添加120 mmol/L乳酸的脑心浸出液肉汤(brain heart infusion broth, BHI) 培养基在厌氧箱内分别培养野生株V. parvula和标准株V. parvula, 检测24 h内野生株V. parvula和标准株V. parvula生长状况及乳酸代谢能力。将野生株V. parvula和标准株V. parvula分别与S. mutans进行共培养, 检测培养基内乳酸浓度及pH值, 用扫描电镜观察细菌生物膜的结构, 结晶紫染色后对细菌生物膜形成量进行半定量分析, 初步探索两者之间的互相作用关系。结果: 经生化鉴定及16S rRNA基因鉴定, 从成年猛性龋患者健康牙面的龈上菌斑中分离出1株野生株V. parvula。24 h内对野生株V. parvula及标准株V. parvula进行培养, 两者生长曲线与培养基内剩余乳酸浓度随时间变化趋势不同, 二者之间的差异有统计学意义(F=10.431, P＜0.001;F=5.641, P＜0.05);将野生株V. parvula和标准株V. parvula分别与S. mutans共培养, 12 h时野生株V. parvula与S. mutans共培养组形成的细菌生物膜结构比标准株V. parvula与S. mutans共培养组更为致密、生物膜量更多(P < 0.001)。24 h时野生株V. parvula与S. mutans共培养组产生的乳酸累积浓度高达65 mmol/L, 显著高于标准株V. parvula与S. mutans共培养组和S. mutans单培养组(P < 0.001)。结论: 利用韦荣氏球菌琼脂培养基能够成功从成年猛性龋患者健康牙面的龈上菌斑中分离野生株V. parvula, 其生长能力、乳酸代谢能力及促进S. mutans形成生物膜和产酸能力均高于标准株V. parvula, 能够加速早期龋的形成。

关键词: 龋病, 猛性龋, 小韦荣球菌, 变异链球菌, 龈上菌斑

Abstract:

Objective: To isolate and cultivate Veillonella parvula (V. parvula) from the supragingival plaque of adult patients intact teeth surfaces with rampant caries, to investigate its growth and metabolic properties, and to preliminarily explore its interaction with Streptococcus mutans (S. mutans). Methods: V. parvula was isolated from the supragingival plaque of intact teeth surfaces in an adult patient with rampant caries using Veillonella agar medium. Identification was performed based on colony morphology, biochemical tests, and 16S rRNA gene sequencing. The clinically isolated strain was cultured in brain heart infusion broth (BHI) supplemented with 120 mmol/L lactate in an anaerobic chamber. Its growth curve and lactate metabolism over 24 h were assessed. Co-culture with S. mutans was conducted to measure lactate accumulation and pH value changes in the culture system. Biofilm structure was observed by scanning electron microscopy (SEM), and the biofilm biomass was compared using crystal violet staining, providing initial insights into their interaction. Results: Through biochemical identification and 16S rRNA gene sequencing, one wild type strain of V. parvula was isolated from the supragingival plaque of intact teeth surfaces in an adult patient with rampant caries. When the wild type V. parvula and the reference strain V. parvula were cultured over 24 h, their growth curves and the trends in the residual lactate concentration in the medium differed, with the differences being statistically significant (F=10.431, P < 0.001; F=5.641, P < 0.05). In co-culture with S. mutans, the group with the wild type V. parvula formed a denser bacterial biofilm structure and had a greater biofilm biomass at 12 h compared with the group with the reference strain V. parvula (P < 0.001). At 24 h, the cumulative lactate concentration produced by the co-culture group with the wild type V. parvula reached as high as 65 mmol/L, which was significantly higher than that in the co-culture group with the reference strain V. parvula and the S. mutans mono-culture group (P < 0.001). Conclusion: The strain of V. parvula which we isolated from supragingival dental plaque exhibited superior growth, lactate metabolism, and a greater capacity to promote S. mutans biofilm formation than the reference strain, ultimately accelerating the initiation of early carious lesions.

Key words: Dental caries, Rampant caries, Veillonella parvula, Streptococcus mutans, Supragingival plaque

中图分类号: 

  • R781.4

图1

临床分离株的菌落形态"

图2

临床分离株的系统发育树"

图3

野生株V. parvula与标准株V. parvula的生长曲线及培养基内乳酸浓度"

图4

扫描电镜下观察生物膜形态结构"

图5

光学显微镜下观察结晶紫染色结果(× 1 000)"

图6

结晶紫染色的半定量分析细菌生物膜量"

图7

V. parvula与S. mutans共培养组中乳酸积累量及pH值的变化"

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ISSN 1671-167X
CN 11-4691/R
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