舌鳞状细胞癌来源的C. sputigena的分离、鉴定及全基因组分析

doi:10.19438/j.cjoms.2024.06.002

中国口腔颌面外科杂志 ›› 2024, Vol. 22 ›› Issue (6): 529-535.doi: 10.19438/j.cjoms.2024.06.002

舌鳞状细胞癌来源的C. sputigena的分离、鉴定及全基因组分析

关智辉^1,2, 李茹³, 于晓芸⁴, 王莉莉⁵, 于苗苗⁶, 巩陆阳¹, 袁荣涛^2,*, 周建华^2,*

1.山东第二医科大学口腔医学院,山东潍坊 261053;
2.青岛市市立医院口腔科,3.医院感染管理科,山东青岛 266071;
4.大连医科大学口腔医学院,辽宁大连 116044;
5.青岛市市立医院中心实验室,山东青岛 266071;
6.滨州医学院口腔医学院,山东烟台 264003

收稿日期:2024-01-21 修回日期:2024-05-08 出版日期:2024-11-20 发布日期:2024-12-11
通讯作者: 袁荣涛,E-mail： yuanrongtao@163.com;
作者简介:关智辉（1997-）,男,在读硕士研究生,E-mail：jx_gzh@163.com
基金资助:
青岛市医药卫生指导计划（2021-WJZD006）

Identification and genome-wide analysis of C. sputigena isolated from squamous cell carcinoma of the tongue

GUAN Zhi-hui^1,2, LI Ru³, YU Xiao-yun⁴, WANG Li-li⁵, YU Miao-miao⁶, GONG Lu-yang¹, YUAN Rong-tao², ZHOU Jian-hua²

1. School of Stomatology, Shandong Second Medical University. Weifang 261053, Shandong Province;
2. Department of Stomatology, 3.Department of Hospital Infection, Qingdao Municipal Hospital. Qingdao 266071, Shandong Province;
4. School of Stomatology, Dalian Medical University. Dalian 116044, Liaoning Province;
5. Central Laboratory, Qingdao Municipal Hospital. Qingdao 266071, Shandong Province;
6. School of Stomatology, Binzhou Medical University. Yantai 264003, Shandong Province, China

Received:2024-01-21 Revised:2024-05-08 Online:2024-11-20 Published:2024-12-11

摘要/Abstract

摘要： 目的： 分离及鉴定舌鳞状细胞癌（tongue squamous cell carcinoma,TSCC）来源的生痰二氧化碳嗜纤维菌（C. sputigena）,从基因组水平分析其耐药和毒力特征。方法： 对TSCC患者的癌组织黏膜的细菌进行分离培养,获得菌株n105及n106。通过菌落形态、革兰染色、16SrRNA序列分析及进化树构建鉴定菌种,采用纸片扩散法检测菌株对4种抗菌药物（克拉霉素、对甲硝唑、青霉素和左氧氟沙星）的敏感性,采用第三代测序技术对菌株n106进行全基因组测序。结果： 菌株n105及n106 16S rRNA序列经BLAST比对及构建Neighbour-Joining进化树,鉴定为C. sputigena。药敏实验表明,C. sputigena菌株n105及n106对克拉霉素敏感,对甲硝唑、青霉素和左氧氟沙星均耐药。全基因组测序结果发现,C. sputigena菌株n106的基因组长度为2.86 Mb,鸟嘌呤-胞嘧啶含量为38.36%,编码2 609个基因。预测毒力基因为tufA、app、clpV、cpsA、acpXL和fHbp,耐药基因为blaCSP-1和erm(F)。结论： TSCC来源的C. sputigena对甲硝唑、青霉素和左氧氟沙星耐药,携带多种毒力及耐药基因,具备潜在致病特征。从微生物角度为早期防控和治疗TSCC提供了新的策略。

关键词: 生痰二氧化碳嗜纤维菌, 舌鳞状细胞癌, 基因组学, 耐药, 毒力基因

Abstract: PURPOSE: To isolate and identify Capnocytophaga sputigena(C. sputigena) from tongue squamous cell carcinoma(TSCC), then analyze the drug resistance and virulence genes at genome-wide level. METHODS: The bacterial strains n105 and n106 were isolated and cultured from the cancer tissue mucosa of TSCC patients. Bacterial species were determined through colony morphology, Gram staining, 16S rRNA sequence, and construction of evolutionary tree. Then the drug sensitivity of bacterial strains to 4 antibiotics(clarithromycin, metronidazole, penicillin and levofloxacin) was detected by disk diffusion method. Whole genome sequencing and functional annotation of virulence and resistance genes were performed on strain n106 by the third-generation sequencing technology. RESULTS: Strains n105 and n106 were identified as C. sputigena through 16S rRNA sequence BLAST alignment and construction of Neighbor Joining evolutionary trees. The results of drug sensitivity test showed that C. sputigena strains n105 and n106 were sensitive to clarithromycin, while resistant to metronidazole, penicillin, and levofloxacin. Whole genome sequencing results showed that the genome length of C. sputigena strain n106 was 2.86 Mb, with 38.36% of guanine cytosine(GC) content, encoding 2 609 genes. Six virulence genes (tufA, app, clpV, cpsA, acpXL and fHbp) and two drug resistance genes [blaCSP-1and erm(F)] were predicted. CONCLUSIONS: C. sputigena isolated from TSCC is resistant to metronidazole, penicillin, and levofloxacin, carrying multiple virulence and resistance genes, which possessing potential pathogenic characteristics. This study provides new strategies for effective prevention, control, and treatment of TSCC from a microbial perspective.

Key words: Capnocytophaga sputigena, Tongue squamous cell carcinoma, Genomics, Drug resistance, Virulence genes

中图分类号:

R739.8

关智辉, 李茹, 于晓芸, 王莉莉, 于苗苗, 巩陆阳, 袁荣涛, 周建华. 舌鳞状细胞癌来源的C. sputigena的分离、鉴定及全基因组分析[J]. 中国口腔颌面外科杂志, 2024, 22(6): 529-535.

GUAN Zhi-hui, LI Ru, YU Xiao-yun, WANG Li-li, YU Miao-miao, GONG Lu-yang, YUAN Rong-tao, ZHOU Jian-hua. Identification and genome-wide analysis of C. sputigena isolated from squamous cell carcinoma of the tongue[J]. China J Oral Maxillofac Surg, 2024, 22(6): 529-535.

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舌鳞状细胞癌来源的C. sputigena的分离、鉴定及全基因组分析

Identification and genome-wide analysis of C. sputigena isolated from squamous cell carcinoma of the tongue

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