上海交通大学学报（医学版）

›› 2012, Vol. 32 ›› Issue (1): 64-.doi: 10.3969/j.issn.1674-8115.2012.01.012

• 论著（临床研究） • 上一篇    下一篇

牙龈卟啉单胞菌临床菌株的分离、鉴定及生长特性分析

李云鹏, 刘大力, 束 蓉, 周彦玢   

  1. 上海交通大学 医学院第九人民医院牙周病科 上海市口腔医学重点实验室, 上海 200011
  • 出版日期:2012-01-28 发布日期:2012-01-29
  • 通讯作者: 束 蓉, 电子信箱: shurong123@hotmail.com。
  • 作者简介:李云鹏 (1985—), 男, 硕士；电子信箱: Inzaghi343882@yahoo.com。
  • 基金资助:

    国家自然科学基金青年基金（30901672）；上海市口腔基础医学重点学科课题 （S30206-fzd03）； 上海市教委科研创新项目(09YZ78)

Isolation, identification and growth characteristics of Porphyromonas gingivalis clinical strains

LI Yun-peng, LIU Da-li, SHU Rong, ZHOU Yan-bin   

  1. Department of Periodontology, the Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai 200011, China
  • Online:2012-01-28 Published:2012-01-29
  • Supported by:

    National Natural Science Foundation of China, 30901672；Foundation of Shanghai Key Basic Discipline of Stomotology, S30206-fzd03；Shanghai Education Committee Foundation, 09YZ78

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摘要：

目的 对口腔中牙龈卟啉单胞菌（P. gingivalis）进行分离和鉴定，初步分析P. gingivalis的生长特性。方法 采集慢性牙周炎患者和牙周健康者临床龈下菌斑标本并行厌氧培养，采用PCR和16S rDNA序列分析方法鉴定P. gingivalis，分析检出率；观察P. gingivalis临床菌株的菌落特征和生长特点，分析生物学特征。结果 从35名采样对象的146个龈下菌斑标本中分离获得6株P. gingivalis临床菌株，分别命名为L2、L3、L4、L5、L11、和L12；牙周健康者龈下菌斑中P. gingivalis的检出率为27.4%，慢性牙周炎患者的牙周健康部位的龈下菌斑中P. gingivalis检出率为86.7%，探诊深度为4~6 mm的牙周袋龈下菌斑中P. gingivalis检出率为95.6%，探诊深度>6 mm的牙周袋龈下菌斑中P. gingivalis检出率为96.0%。L2、L5、L11和L12在BHI平板上培养1周后均形成特征性黑色菌落，而L3和L4在培养1周后形成灰色菌落，10 d后形成特征性黑色菌落；L2、L4、L5和L12在传代30 h后进入对数期生长期，L3和L11则在传代60 h后进入对数生长期。结论 P. gingivalis是慢性牙周炎患者的优势致病菌之一，其在慢性牙周炎患者健康位点的检出率显著高于牙周健康者，且随着牙周探诊深度的加深，检出率呈现逐渐升高的趋势。P. gingivalis各临床菌株的生长存在差异，提示其生物学特征和致病性可能不同。

关键词: 牙龈卟啉单胞菌, 临床菌株, PCR, 生长特性

Abstract:

Objective To isolate and identify Porphyromonas gingivalis (P. gingivalis) from clinical subjects, and analyse the growth characteristics of P. gingivalis. Methods Subgingival plaque samples were collected from patients with chronic periodontitis and periodontally healthy subjects, and anaerobic culture was performed. P. gingivalis strains were identified by PCR and 16S rDNA sequencing, and detection rate was analysed. Bacteria colonies and growth characteristics of the clinical strains of P. gingivalis were observed, and the biological properties were analysed. Results Six P. gingivalis clinical strains named as L2, L3, L4, L5, L11 and L12 were isolated from 146 subgingival plaque samples collected from 35 subjects. The detection rate of P. gingivalis was 27.4% in subgingival plaque samples of periodontally healthy subjects, 86.7% in periodontally healthy sites of patients with chronic periodontitis, 95.6% in periodontal pockets with probing depth of 4-6 mm, and 96.0% in periodontal pockets with probing depth >6 mm. Strains L2, L5, L11 and L12 formed black colonies on BHI plates after culture for 1 week. Strains L3 and L4 formed grey colonies 1 week after culture for 1 week, and formed black colonies after culture for 10 d. Strains L2, L4, L5 and L12 grew to exponential phase 30 h after inoculation, whereas strains L3 and L11 grew to exponential phase 60 h after inoculation. Conclusion P. gingivalis is one of the dominant pathogenic bacteria in periodontal pockets of patients with chronic periodontitis. The detection rate of P. gingivalis in healthy sites of patients with chronic periodontitis is significantly higher than that of periodontally healthy subjects, and the detection rates of P. gingivalis are higher in deeper periodontal pockets. The growth characteristics of P. gingivalis clinical strains vary from each other, indicating that the biological and pathogenic properties of P. gingivalis may be strain-specific.

Key words: Porphyromonas gingivalis, clinical strains, PCR, growth characteristics