Mi Mengheng, Wu Fangcao, Zhu Jian, Liu Fang, Cui Guzhen, Wen Xueqing, Hu Yue, Deng Zhaohui, Wu Xiaojuan, Zhang Zhengrong, Qi Tingna, Chen Zhenghong
Key Laboratory of Microbiology and Parasitology of Education Department of Guizhou, School of Basic Medical Science, Guizhou Medical University/Joint Laboratory of Helicobacter Pylori and Intestinal Microecology of Affiliated Hospital of Guizhou Medical University, Guiyang, People's Republic of China.
Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University, Guiyang, People's Republic of China.
Infect Drug Resist. 2021 Feb 11;14:535-545. doi: 10.2147/IDR.S287631. eCollection 2021.
Chronic infection causes peptic ulcers in a subpopulation of individuals and is a risk factor for the development of gastric cancer. Multiple infections and heteroresistant contribute to poor treatment efficacy. Here, we investigated the extent of genetic diversity among strains within a given host and its influence on the results of antibiotic (metronidazole, levofloxacin, clarithromycin, amoxicillin, and tetracycline) susceptibility testing.
Gastric mucosa biopsy samples were obtained from patients with gastric disorders, including 48 positive patients, who were never previously treated for infection. Five potential colonies isolated from each sample were subcultured for enrichment. Enriched colonies were identified through Gram staining and assays for urease, oxidase, and catalase. For each monoclonal colony, the antibiotic susceptibility was assessed, genomic DNA was sequenced, and the () genotype was verified. Co-infection with multiple strains was determined using random amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR).
Thirteen gastric mucosa biopsy samples were positive for . Five monoclonal strains isolated from each of these 13 patients were identified as . RAPD-PCR indicated that intra-patient monoclonal strains of in 10 of the 13 samples exhibited heterogeneity. Among the 13 patients, intra-patient monoclonal strains isolated from 4 patients had identical genotype, whereas intra-patient monoclonal strains isolated from the other 9 patients harbored more than one genotype. The antibiotic susceptibility of five intra-patient monoclonal strains from seven patients was inconsistent.
The existence of heterogeneous strains with resistance to different drugs and virulence were common within the gastric mucosa of an individual patient.
慢性感染在部分个体中会导致消化性溃疡,并且是胃癌发生的一个危险因素。多种感染及异质性耐药会导致治疗效果不佳。在此,我们研究了给定宿主内菌株的基因多样性程度及其对抗生素(甲硝唑、左氧氟沙星、克拉霉素、阿莫西林和四环素)药敏试验结果的影响。
从患有胃部疾病的患者获取胃黏膜活检样本,其中包括48例幽门螺杆菌阳性患者,这些患者此前从未接受过幽门螺杆菌感染治疗。从每个样本中分离出的5个潜在幽门螺杆菌菌落进行传代培养以富集。通过革兰氏染色以及脲酶、氧化酶和过氧化氢酶检测对富集后的幽门螺杆菌菌落进行鉴定。对于每个幽门螺杆菌单克隆菌落,评估其抗生素敏感性,对基因组DNA进行测序,并验证幽门螺杆菌(Hp)基因型。使用随机扩增多态性DNA(RAPD)-聚合酶链反应(PCR)确定是否存在多种幽门螺杆菌菌株共感染。
13份胃黏膜活检样本幽门螺杆菌呈阳性。从这13例患者中的每例分离出的5个单克隆菌株均鉴定为幽门螺杆菌。RAPD-PCR表明,13个样本中有10个样本的患者体内幽门螺杆菌单克隆菌株表现出异质性。在这13例患者中,从4例患者分离出的患者体内幽门螺杆菌单克隆菌株具有相同的幽门螺杆菌基因型,而从其他9例患者分离出的患者体内幽门螺杆菌单克隆菌株含有不止一种幽门螺杆菌基因型。7例患者的5个患者体内幽门螺杆菌单克隆菌株的抗生素敏感性不一致。
对不同药物具有耐药性和毒力的异质性幽门螺杆菌菌株在个体患者的胃黏膜中普遍存在。