胃黏膜组织中恶臭假单胞菌临床意义的探讨

发布时间：2018-11-08 18:36

【摘要】：目的探索恶臭假单胞菌在幽门螺杆菌感染相关胃肠疾病中的意义。方法采集14 C尿素呼气试验阳性患者病变胃黏膜354例,进行细菌的分离培养。根据菌落形态、革兰染色、尿素酶试验及幽门螺杆菌特异性16SrRNA基因片段的PCR进行幽门螺杆菌的鉴定,同时提取胃黏膜组织DNA,通过幽门螺杆菌特异性PCR进行快速诊断。将非幽门螺杆菌转种于营养琼脂培养基,进行快速尿素酶试验。尿素酶阳性的非幽门螺杆菌染色体DNA利用细菌16SrRNA基因通用引物进行PCR扩增、测序及序列比对。使用全自动细菌鉴定仪对经测序比对为恶臭假单胞菌的菌株进行鉴定。采用K-B纸片扩散法进行药物敏感性试验。结果从354例样本中分离出革兰阴性、尿素酶阳性的恶臭假单胞菌10株,经16SrRNA基因测序和序列比对,与GenBank中恶臭假单胞菌的相似性≥98%。10株恶臭假单胞菌的胃黏膜标本中,6例标本幽门螺杆菌特异性PCR为阳性,4例为阴性。传代存活的7株恶臭假单胞菌的K-B法药物敏感试验显示对左氧氟沙星均敏感,1株对四环素敏感,5株对阿莫西林耐药,6株对克拉霉素耐药,7株对甲硝唑、氨苄青霉素均耐药。7株菌经全自动细菌生化鉴定仪鉴定结果均为恶臭假单胞菌。结论病变胃黏膜中分离出尿素酶阳性且对治疗幽门螺杆菌感染的多种抗生素耐药;可能造成临床上14 C-尿素呼气试验假阳性,并继发感染影响胃肠疾病的进展。
[Abstract]:Objective to explore the significance of Pseudomonas odour in Helicobacter pylori infection related gastrointestinal diseases. Methods 354 patients with 14 C urea breath test positive gastric mucosa were collected for bacterial isolation and culture. Helicobacter pylori was identified by colony morphology, Gram staining, urease test and PCR of Helicobacter pylori specific 16SrRNA gene fragment. DNA, was extracted from gastric mucosa for rapid diagnosis by Helicobacter pylori specific PCR. Non-Helicobacter pylori was transferred to nutrient Agar medium for rapid urease test. Urea-positive non-Helicobacter pylori chromosome DNA was amplified by PCR, sequenced and sequenced using universal primers of bacterial 16SrRNA gene. A fully automatic bacterial identification instrument was used to identify the strains of Pseudomonas odour by sequencing. Drug sensitivity test was carried out by K-B disk diffusion method. Results Ten strains of gram-negative and urease positive Pseudomonas oxysporum were isolated from 354 samples. By sequencing and sequencing of 16SrRNA gene, the similarity of 10 strains of Pseudomonas oxysporum in GenBank was more than 98%. Helicobacter pylori specific PCR was positive in 6 cases and negative in 4 cases. K-B susceptibility test of 7 strains of Pseudomonas oxysporum by subculture showed that all strains were sensitive to levofloxacin, 1 strain was sensitive to tetracycline, 5 strains were resistant to amoxicillin, 6 strains were resistant to clarithromycin, 7 strains were resistant to metronidazole, 5 strains were resistant to amoxicillin, 6 strains were resistant to clarithromycin, 7 strains were resistant to metronidazole. All of the 7 strains were resistant to ampicillin. Conclusion Urease positive and antibiotic resistance in the treatment of Helicobacter pylori infection may result in false positive 14C-urea breath test and secondary infection may affect the progression of gastrointestinal diseases.
【作者单位】：贵州医科大学医学检验学院临床生化教研室;贵州医科大学基础医学院微生物学教研室;贵州医科大学附属医院消化内科;黔南布依族苗族自治州人民医院消化内科;贵阳市儿童医院消化内科;
【基金】：国家自然科学基金资助项目(No.81460314) 贵阳市卫生和计划生育委员会科学技术计划项目([2014]筑卫计科技合同字第018号)联合资助~~
【分类号】：R57

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