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炎症性肠病患者肠道脆弱拟杆菌菌株中脆弱拟杆菌群的检测以及bft肠毒素和抗生素耐药标记物cepA、cfiA和nim的多样性

Detection of B. fragilis group and diversity of bft enterotoxin and antibiotic resistance markers cepA, cfiA and nim among intestinal Bacteroides fragilis strains in patients with inflammatory bowel disease.

作者信息

Rashidan Marjan, Azimirad Masoumeh, Alebouyeh Masoud, Ghobakhlou Mehdi, Asadzadeh Aghdaei Hamid, Zali Mohammad Reza

机构信息

Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

出版信息

Anaerobe. 2018 Apr;50:93-100. doi: 10.1016/j.anaerobe.2018.02.005. Epub 2018 Feb 14.

Abstract

We compared frequency of the members of B. fragilis group in 100 and 20 colon biopsy specimens of inflammatory bowel disease (IBD) and non-IBD patients. Agar dilution and PCR were orderly used to detect minimal inhibitory concentration of ampicillin, imipenem, and metronidazole, and carriage of related resistance genes cepA, cfi, and nim. B. fragilis group was detected in 38% of IBD (UC: 36/89; CD:1/11) and 25% (5/20) of non-IBD patients. While B. vulgatus (UC: 20/36, CD: 1/2, control: 1/6); B. fragilis (UC: 18/36, CD: 1/2, control: 5/6); B. ovatus (UC: 2/36); B. caccae (UC: 1/36); and B. eggerthii (UC: 1/36) were characterized, colonization of B. thetaiotamicron, B. merdae, B. distasonis, B. stercoris and B. dorei species was not detected in these specimens. Co-existence of B. fragilis + B. vulgatus (5 patients) and B. vulgatus + B. caccae (1 patient) was detected just in UC patients. bft was detected among 31.5% (6/19) of B. fragilis strains in the IBD and 40% (2/5) in the non-IBD groups. Nearly, 73.6% of the strains from the patient group and 80% in control group harbored cepA; 31.5% and 20% in the patients and control groups harbored cfiA, and none of them harbored nim determinant. Co-occurrence of the cepA and cfiA was orderly detected in 10.5% (2/19) and 20% (1/5) of the strains in these groups. The resistance rates were detected as 95.8% (23/24 (to ampicillin (MIC range of ≤0.5-≥16 μg/ml), 0% to metronidazole and 29.1% to imipenem (7/24, MIC range ≤4-32 μg/ml). Nearly 25% (6/24) of the strains were resistant to ampicillin and imipenem, simultaneously. No statistically significant difference was detected between the IBD and control groups for drug resistance phenotypes. Statistical analysis showed significant associations between resistance to ampicillin or imipenem and carriage of cepA or cfiA, respectively (p value = 0.0007). PCR results on the extracted plasmids confirmed their roles in carriage of cfiA and cepA. These data provide guide for antibiotic therapy and highlights wide distribution of β-lactam resistant B. fragilis strains in patients with IBD and non-IBD intestinal disorders.

摘要

我们比较了脆弱拟杆菌群成员在100例炎症性肠病(IBD)患者和20例非IBD患者的结肠活检标本以及100例IBD患者和20例非IBD患者的结肠活检标本中的频率。依次采用琼脂稀释法和PCR检测氨苄西林、亚胺培南和甲硝唑的最低抑菌浓度,以及相关耐药基因cepA、cfi和nim的携带情况。在38%的IBD患者(溃疡性结肠炎:36/89;克罗恩病:1/11)和25%(5/20)的非IBD患者中检测到脆弱拟杆菌群。鉴定出了普通拟杆菌(溃疡性结肠炎:20/36,克罗恩病:1/2,对照组:1/6)、脆弱拟杆菌(溃疡性结肠炎:18/36,克罗恩病:1/2,对照组:5/6)、卵形拟杆菌(溃疡性结肠炎:2/36)、粪便拟杆菌(溃疡性结肠炎:1/36)和埃氏拟杆菌(溃疡性结肠炎:1/36),在这些标本中未检测到多形拟杆菌、屎拟杆菌、解脲拟杆菌、硬结拟杆菌和多雷拟杆菌的定植。仅在溃疡性结肠炎患者中检测到脆弱拟杆菌+普通拟杆菌共存(5例患者)和普通拟杆菌+粪便拟杆菌共存(1例患者)。在IBD组31.5%(6/19)的脆弱拟杆菌菌株和非IBD组40%(2/5)的脆弱拟杆菌菌株中检测到脆弱拟杆菌毒素。患者组近73.6%的菌株和对照组80%的菌株携带cepA;患者组和对照组分别有31.5%和20%的菌株携带cfiA,且均未携带nim决定簇。在这些组中,依次在10.5%(2/19)和20%(1/5)的菌株中检测到cepA和cfiA的共出现。耐药率检测为95.8%(23/24,对氨苄西林(MIC范围≤0.5 - ≥16μg/ml))、对甲硝唑0%、对亚胺培南29.1%(7/24;MIC范围≤4 - 32μg/ml)。近25%(6/24)的菌株同时对氨苄西林和亚胺培南耐药。IBD组和对照组在耐药表型上未检测到统计学显著差异。统计分析显示,对氨苄西林或亚胺培南耐药分别与携带cepA或cfiA之间存在显著关联(p值 = 0.0007)。对提取质粒的PCR结果证实了它们在携带cfiA和cepA中的作用。这些数据为抗生素治疗提供了指导,并突出了β-内酰胺耐药脆弱拟杆菌菌株在IBD和非IBD肠道疾病患者中的广泛分布。

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