摘要
目的筛选一种适合牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)生长的固体琼脂培养基,建立能有效、快速分离Pg菌株的方法。方法通过观察比较Pg标准菌株ATCC 33277在苛养厌氧琼脂平板(FAA)、胰酪大豆胨琼脂平板(TSA)和牛脑心浸液琼脂平板(BHIA)上的生长周期和菌落特点,筛选最佳平板。采集食管癌高发地区1例慢性牙周炎患者牙菌斑,并收集3例食管鳞癌患者的新鲜肿瘤组织,经实时荧光定量PCR(qPCR)鉴定Pg为阳性,在FAA平板上连续划线进行常规厌氧培养。采用革兰氏染色、qPCR和16S rRNA测序等方法初步鉴定分离的临床菌株,并开展生长曲线、药物敏感性及生物膜形成能力测试等实验,进一步研究分离临床菌株的生物学特性。结果与BHIA和TSA相比,FAA更适合Pg的生长,其灵敏性更高。从口腔中成功分离一株Pg临床菌株,命名为LyG1;食管鳞癌组织中分离一株Pg临床菌株,命名为LyEC01。结论FAA适合分离、纯化及培养Pg,利用FAA从口腔和食管癌组织中分离出的LyG1和LyEC01临床菌株,与标准菌株的生物学特性存在差异。
Objective To select a solid agar medium suitable for the growth of Porphyromonas gingivalis(Pg)and to establish an effective and rapid method for the isolation of Pg strains.Methods By observing the growth cycle and colony characteristics of Pg,we investigated the growth characteristics of Pg standard strain ATCC 33277 on fastidious anaerobic agar(FAA),tryptic soy peptone agar(TSA),and bovine brain heart infusion agar(BHIA).BHIA were used to characterize the growth and screen the best plates.Dental plaque was collected from 1 patient with chronic periodontitis in an area with high incidence of esophageal cancer,and fresh tumor tissue from 3 patients with esophageal squamous cell carcinoma was collected,identi-fied as positive for Pg by real-time fluorescence quantitative PCR(qPCR),and cultured on FAA plates with continuous lines for routine anaerobic culture.Gram staining,qPCR and 16S rRNA sequencing were used to preliminarily identify the isolated clinical strains,and experiments such as growth curve test,drug sensitivity and biofilm-forming ability were carried out to further study the biological characteristics of the isolated clinical strains.Results Compared with BHIA and TSA,FAA was more suitable for the growth of Pg and its sensitivity was higher.A clinical strain of Pg,named LyG1,was successfully isolated from oral cavity,and a clinical strain of Pg,named LyEC01,was isolated from esophageal squamous carcinoma tissues.Conclusion FAA is suitable for the isolation,purification as well as cultivation of Pg,and the clinical strains of LyG1 and LyEC01,which were isolated from oral cavity and esophageal carcinoma tissues,differed from the standard strains in their biological characteristics.
作者
杨睿
裴广坤
石林林
左志向
颜贞
谷变利
高社干
YANG Rui;PEI Guangkun;SHI Linlin;ZUO Zhixiang;YAN Zhen;GU Bianli;GAO Shegan(Henan Key Laboratory of Microbiome and Esophageal Cancer Prevention and Treatment,Henan Key Laboratory of Cancer Epigenetics,Cancer Hospital,The First Affiliated Hospital(College of Clinical Medicine)of Henan University of Science and Technology,Luoyang,China,471003;Sun Yat-sen University Cancer Center,Guangzhou,China,510060;Jiangsu Hengrui Medicine Co.,Ltd.,Lianyungang,China,222000)
出处
《食管疾病》
2025年第2期81-87,共7页
Journal of Esophageal Diseases
基金
河南省重点研发与推广专项(科技攻关)项目(222102310115)河南省医学科技攻关省部共建重点项目(SBGJ202102199)。
关键词
食管癌
牙龈卟啉单胞菌
苛养厌氧平板
临床菌株
esophageal cancer
Porphyromonas gingivalis
fastidious anaerobic agar
clinical strains
