幽门螺杆菌毒力基因型与儿童胃十二指肠疾病关系以及毒力基因对根除治疗的影响

发布时间：2018-04-18 23:30

本文选题：儿童 + 幽门螺杆菌　；参考：《南昌大学》2016年博士论文

【摘要】：背景和目的:H.pylori感染与儿童多种上消化道疾病密切相关。H.pylori毒力基因主要有cag A,vac A和ice A基因。H.pylori感染定植在胃黏膜后,导致一系列疾病的发生。随着儿童感染H.pylori根除治疗三联疗法的应用,临床出现了抗生素的耐药菌株。研究表明,H.pylori毒力基因型和抗生素耐药菌株的出现是影响H.pylori根除治疗效果的两个重要因素。本课题对江西地区患儿胃粘膜进行H.pylori分离培养和药敏实验,检测cag A,vac A及ice A基因型,H.pylori感染患儿采用三联疗法根除治疗,以明确江西地区H.pylori菌株培养情况和抗生素的耐药性,H.pylori毒力基因的优势基因型,研究H.pylori毒力基因型与儿童胃十二指肠疾病和耐药性之间的关系,以及毒力基因型对根除治疗的影响。材料和方法:1、研究对象2014年7月~2015年8月因具有上消化道症状在江西省儿童医院门诊就诊和住院的患儿,共316例纳入本研究,并进行胃镜检查和胃窦黏膜活检。2、H.pylori分离培养和药敏实验胃黏膜活检标本研磨后涂布于培养基表面,放置于三气混合恒温孵养箱内培养获得H.pylori阳性菌株,然后采用E-test法检测阿莫西林、克拉霉素和甲硝唑的最低抑菌浓度(MIC);利用K-B法检测呋喃唑酮的敏感性。3、H.pylori毒力基因PCR扩增H.pylori菌株用QIAamp DNA mini kit试剂盒抽提H.pylori基因组,使用Nano Drop 2000超微量分光光度计检测所提取菌株基因组DNA浓度。根据文献中H.pylori毒力基因的引物序列进行PCR扩增,采用琼脂糖凝胶电泳后,使用Bio-Rad Gel Doc XR+凝胶成像分析仪进行图像分析。4、H.pylori感染患儿采用质子泵抑制剂加两种抗生素的三联疗法根除治疗。结果:1、H.pylori分离培养情况及耐药性分析在316例患儿胃窦黏膜共培养出107株H.pylori菌株,检出率33.9%,其中男性81例、女性26例,年龄分布1岁9个月至14岁。在慢性胃炎、十二指肠球炎、胃溃疡和十二指肠球部溃疡4组中H.pylori分离培养阳性检出率分别为20.1%,44.8%,30.0%和52.8%(χ2=33.133,P0.001);其中十二指肠球部溃疡阳性检出率高于慢性胃炎和胃溃疡组(χ2=31.861,P0.001;χ2=1.098,P=0.295),十二指肠球炎阳性率检出高于慢性胃炎和胃溃疡组(χ2=8.348,P=0.004;P=0.480)。在107株菌株中,97株完成药敏实验,对克拉霉素耐药率20.6%,对甲硝唑耐药率47.4%,对克拉霉素+甲硝唑二重耐药率12.4%,未发现对阿莫西林和呋喃唑酮耐药菌株;H.pylori对克拉霉素和甲硝唑2组间耐药率的比较差异有统计意义(χ2=15.524,P0.001)。2、H.pylori毒力基因型与儿童胃十二指肠疾病之间的关系在107株菌株中,cag A基因阳性检出率为94.4%。在vac A基因亚型中,vac As1a和vac Am2基因亚型阳性检出率最高,分别74.8%和69.2%,0.9%菌株同时检测出vac Am1和vac Am2基因亚型;在vac A基因的嵌合体中,vac As1a/m2基因亚型阳性检出率最高,51.4%(χ2=69.283,P0.001);但是在所有菌株中未检测出vac Aslb和vac As2基因亚型。ice A1和ice A2基因亚型单独阳性检出率分别为79.4%和9.3%(χ2=113.822,P0.001),其中7.5%菌株同时检出ice A1和ice A2基因亚型。H.pylori毒力基因型在消化性溃疡、慢性胃炎和十二指肠球炎3组间的阳性检出率比较差异无统计学意义(P0.05);cag A/vac As1a/m2基因组合型在消化性溃疡、慢性胃炎和十二指肠球炎3组间的阳性检出率比较,差异无统计学意义(χ2=2.670,P=0.263)。3、H.pylori毒力基因型与抗生素耐药性之间的关系H.pylori毒力基因在克拉霉素耐药组、甲硝唑耐药组、克拉霉素+甲硝唑二重耐药组和抗生素敏感组4组间的阳性检出率比较,差异均无统计学意义(P0.05)。4、H.pylori毒力基因型与根除治疗的关系在107例患儿中,98例完成本研究治疗观察,9例失访。H.pylori根除率按意向治疗分析(ITT)及试验方案分析(PP)分别为64.5%和70.4%。H.pylori毒力基因型在根除治疗成功组和根除治疗失败组的阳性检出率的比较差异无统计学意义(p0.05)。结论:1、H.pylori毒力基因型与儿童胃十二指肠疾病类型和耐药无相关性;2、江西地区儿童感染H.pylori优势基因亚型是cag A,vac As1a/m2和ice A1,H.pylori感染存在不同基因亚型菌株混合感染;3、江西地区儿童感染H.pylori对甲硝唑和克拉霉素的耐药率较高并有较高的双重耐药,未检测出对阿莫西林和呋喃唑酮耐药菌株;4、H.pylori cag A,vac A和ice A基因型与以奥美拉唑加两种抗生素组成的14天三联疗法根除疗效无相关性。
[Abstract]:Background and objective: H.pylori infection in children with upper gastrointestinal disease is closely related to a variety of.H.pylori virulence genes are CAG A, VAC A and ice A gene.H.pylori colonization in gastric mucosa, leading to a series of diseases. With the eradication of H.pylori infection in children treated by triple therapy, clinical antibiotic resistant strains. Show that the appearance of H.pylori virulence genotypes and antibiotic resistant strains are two important factors that influence the therapeutic effect of H.pylori eradication. Isolation and drug sensitivity test of H.pylori of children in Jiangxi area of the gastric mucosa, detection of CAG A, VAC A and ice A genotype H.pylori infection were treated with triple eradication therapy, in order to clear resistance in Jiangxi H.pylori strains and antibiotics, the dominant genotype of H.pylori virulence genes, H.pylori genotype and virulence of the stomach in twelve children The relationship between intestinal diseases and drug resistance, and virulence genotype on the eradication effect. Materials and methods: 1, July 2014 ~2015 year in August for the research object with upper gastrointestinal symptoms in outpatients of Jiangxi province children's Hospital and hospital patients, a total of 316 patients enrolled in the study, and gastroscopy and gastric mucosa biopsy.2, H.pylori were isolated and drug sensitive test of gastric mucosal biopsy specimens were obtained after grinding is coated on the surface of the medium, placed in three mixed gas temperature in the incubator incubation of culture H.pylori positive strains, and E-test method was used to detect the minimal inhibitory concentration of amoxicillin, clarithromycin and metronidazole (MIC); the sensitivity of.3 detection of furazolidone by K-B method the H.pylori gene was amplified by PCR QIAamp DNA Mini Kit H.pylori strain H.pylori genome extraction kit, use Nano Drop 2000 ultra micro spectrophotometer detection by extracting bacteria The genomic DNA was amplified by PCR. The concentration of primers according to the sequence in the literature H.pylori virulence gene, by agarose gel electrophoresis, image analysis was performed using Bio-Rad.4 Gel Doc XR+ gel imaging analyzer, triple therapy for H.pylori infection were treated with proton pump inhibitors plus two antibiotics eradication therapy. Results: 1, isolation and drug resistance analysis in 316 cases of gastric mucosa were isolated from 107 strains of H.pylori H.pylori strains, the detection rate of 33.9%, of which 81 were male, 26 were female, the age distribution of 1 years and 9 months to 14 years old. In chronic gastritis, twelve duodenal bulb inflammation, gastric ulcer and duodenal ulcer in 4 groups of H.pylori were isolated and cultured positive rates were 20.1%, 44.8%, 30% and 52.8% (2=33.133, P0.001); the positive rate was higher than that of duodenal ulcer and chronic gastritis and gastric ulcer group (x 2=31.861, P0.001 X; 2=1.098, P=0.295), The positive rate of duodenal inflammation detection is higher than that of chronic gastritis and gastric ulcer group (x 2=8.348, P=0.004; P=0.480). Among the 107 isolates, 97 strains of complete drug sensitivity test, 20.6% for clarithromycin resistance rate, 47.4% to 12.4% of the rate of resistance to metronidazole, clarithromycin and metronidazole resistance rate was found to double, amoxicillin and furazolidone drug resistant strains; there was significant difference between the H.pylori of clarithromycin and metronidazole resistance rate between the 2 groups (2=15.524, P0.001).2, H.pylori virulence genotypes and gastroduodenal disease in children in 107 strains, CAG A gene positive rate of 94.4%. in VAC genotype A, VAC and As1a VAC Am2 gene subtype positive rate is highest, respectively 74.8% and 69.2%, 0.9% strains detected VAC Am1 and VAC Am2 gene subtype; in chimeric VAC A gene in VAC As1a/m2 subtype positive detection rate (51.4% X 2=69.283, P0.001); but in all of the strains were not detected in the VAC Aslb and VAC As2 gene subtype.Ice A1 and ice A2 genotype alone positive rates were 79.4% and 9.3% (2=113.822, P0.001), of which 7.5% strains were ice A1 and ice A2 gene subtype.H.pylori virulence genotypes in peptic ulcer, chronic gastritis and duodenal inflammation between the 3 groups in the positive rate had no significant difference (P0.05); CAG A/vac As1a/m2 genotype in peptic ulcer, chronic gastritis and duodenal inflammation between the 3 groups in the positive rate, the difference was not statistically significant (x 2=2.670,.3, P=0.263) H.pylori type of antibiotic resistance and virulence genes of the relationship between H.pylori gene in clarithromycin resistance group, metronidazole resistant group, clarithromycin and metronidazole resistance group and antibiotic sensitive double positive group between the 4 groups in the detection rate of comparison, difference There were no significant differences (P0.05).4, H.pylori genotype and virulence eradication in 107 patients, 98 patients completed the study treatment, 9 patients lost the eradication rate of.H.pylori according to the intention to treat analysis (ITT) analysis scheme and test (PP) were 64.5% and 70.4%.H.pylori virulence genotypes in successful eradication treatment group and eradication failure group positive rate showed no significant difference (P0.05). Conclusion: 1. There is no correlation between H.pylori gene and gastroduodenal disease in children and the type of drug resistance; 2, Jiangxi area children infected with H.pylori subtype CAG is dominant gene A, VAC As1a/m2 and ice A1, H.pylori there are different infection subtype strains mixed infection; 3, double resistance H.pylori resistance to metronidazole and clarithromycin was higher and higher in children in Jiangxi did not detect the infection of amoxicillin and furazolidone resistant The strains; 4, H.pylori CAG A, VAC A and ice A genotypes were not associated with the 14 day triple therapy, consisting of Omeprazole plus two antibiotics.

【学位授予单位】：南昌大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R725.7

【相似文献】