本文選題：兒童 + 幽門螺桿菌��； 參考：《南昌大學》2016年博士論文

【摘要】：背景和目的:H.pylori感染與兒童多種上消化道疾病密切相關。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,導致一系列疾病的發(fā)生。隨著兒童感染H.pylori根除治療三聯(lián)療法的應用,臨床出現(xiàn)了抗生素的耐藥菌株。研究表明,H.pylori毒力基因型和抗生素耐藥菌株的出現(xiàn)是影響H.pylori根除治療效果的兩個重要因素。本課題對江西地區(qū)患兒胃粘膜進行H.pylori分離培養(yǎng)和藥敏實驗,檢測cag A,vac A及ice A基因型,H.pylori感染患兒采用三聯(lián)療法根除治療,以明確江西地區(qū)H.pylori菌株培養(yǎng)情況和抗生素的耐藥性,H.pylori毒力基因的優(yōu)勢基因型,研究H.pylori毒力基因型與兒童胃十二指腸疾病和耐藥性之間的關系,以及毒力基因型對根除治療的影響。材料和方法:1、研究對象2014年7月~2015年8月因具有上消化道癥狀在江西省兒童醫(yī)院門診就診和住院的患兒,共316例納入本研究,并進行胃鏡檢查和胃竇黏膜活檢。2、H.pylori分離培養(yǎng)和藥敏實驗胃黏膜活檢標本研磨后涂布于培養(yǎng)基表面,放置于三氣混合恒溫孵養(yǎng)箱內培養(yǎng)獲得H.pylori陽性菌株,然后采用E-test法檢測阿莫西林、克拉霉素和甲硝唑的最低抑菌濃度(MIC);利用K-B法檢測呋喃唑酮的敏感性。3、H.pylori毒力基因PCR擴增H.pylori菌株用QIAamp DNA mini kit試劑盒抽提H.pylori基因組,使用Nano Drop 2000超微量分光光度計檢測所提取菌株基因組DNA濃度。根據(jù)文獻中H.pylori毒力基因的引物序列進行PCR擴增,采用瓊脂糖凝膠電泳后,使用Bio-Rad Gel Doc XR+凝膠成像分析儀進行圖像分析。4、H.pylori感染患兒采用質子泵抑制劑加兩種抗生素的三聯(lián)療法根除治療。結果:1、H.pylori分離培養(yǎng)情況及耐藥性分析在316例患兒胃竇黏膜共培養(yǎng)出107株H.pylori菌株,檢出率33.9%,其中男性81例、女性26例,年齡分布1歲9個月至14歲。在慢性胃炎、十二指腸球炎、胃潰瘍和十二指腸球部潰瘍4組中H.pylori分離培養(yǎng)陽性檢出率分別為20.1%,44.8%,30.0%和52.8%(χ2=33.133,P0.001);其中十二指腸球部潰瘍陽性檢出率高于慢性胃炎和胃潰瘍組(χ2=31.861,P0.001;χ2=1.098,P=0.295),十二指腸球炎陽性率檢出高于慢性胃炎和胃潰瘍組(χ2=8.348,P=0.004;P=0.480)。在107株菌株中,97株完成藥敏實驗,對克拉霉素耐藥率20.6%,對甲硝唑耐藥率47.4%,對克拉霉素+甲硝唑二重耐藥率12.4%,未發(fā)現(xiàn)對阿莫西林和呋喃唑酮耐藥菌株;H.pylori對克拉霉素和甲硝唑2組間耐藥率的比較差異有統(tǒng)計意義(χ2=15.524,P0.001)。2、H.pylori毒力基因型與兒童胃十二指腸疾病之間的關系在107株菌株中,cag A基因陽性檢出率為94.4%。在vac A基因亞型中,vac As1a和vac Am2基因亞型陽性檢出率最高,分別74.8%和69.2%,0.9%菌株同時檢測出vac Am1和vac Am2基因亞型;在vac A基因的嵌合體中,vac As1a/m2基因亞型陽性檢出率最高,51.4%(χ2=69.283,P0.001);但是在所有菌株中未檢測出vac Aslb和vac As2基因亞型。ice A1和ice A2基因亞型單獨陽性檢出率分別為79.4%和9.3%(χ2=113.822,P0.001),其中7.5%菌株同時檢出ice A1和ice A2基因亞型。H.pylori毒力基因型在消化性潰瘍、慢性胃炎和十二指腸球炎3組間的陽性檢出率比較差異無統(tǒng)計學意義(P0.05);cag A/vac As1a/m2基因組合型在消化性潰瘍、慢性胃炎和十二指腸球炎3組間的陽性檢出率比較,差異無統(tǒng)計學意義(χ2=2.670,P=0.263)。3、H.pylori毒力基因型與抗生素耐藥性之間的關系H.pylori毒力基因在克拉霉素耐藥組、甲硝唑耐藥組、克拉霉素+甲硝唑二重耐藥組和抗生素敏感組4組間的陽性檢出率比較,差異均無統(tǒng)計學意義(P0.05)。4、H.pylori毒力基因型與根除治療的關系在107例患兒中,98例完成本研究治療觀察,9例失訪。H.pylori根除率按意向治療分析(ITT)及試驗方案分析(PP)分別為64.5%和70.4%。H.pylori毒力基因型在根除治療成功組和根除治療失敗組的陽性檢出率的比較差異無統(tǒng)計學意義(p0.05)。結論:1、H.pylori毒力基因型與兒童胃十二指腸疾病類型和耐藥無相關性;2、江西地區(qū)兒童感染H.pylori優(yōu)勢基因亞型是cag A,vac As1a/m2和ice A1,H.pylori感染存在不同基因亞型菌株混合感染;3、江西地區(qū)兒童感染H.pylori對甲硝唑和克拉霉素的耐藥率較高并有較高的雙重耐藥,未檢測出對阿莫西林和呋喃唑酮耐藥菌株;4、H.pylori cag A,vac A和ice A基因型與以奧美拉唑加兩種抗生素組成的14天三聯(lián)療法根除療效無相關性。
[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.

【學位授予單位】：南昌大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：R725.7

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