广州地区患儿感染幽门螺杆菌vacA、cagA及iceA基因研究

目的探讨广州地区患儿感染幽门螺杆菌（Hp）的vacA、cagA、iceA的基因亚型和优势基因亚型。方法105例胃、十二指肠疾病患儿的胃窦处取3块胃黏膜,分别进行快速尿素酶反应、病理检查和聚合酶链反应（PCR）。抽提胃黏膜基因组DNA,用11对引物,检测HpureA、vacA、cagA和ieeA基因,分析HpvacA、cagA、iceA基因亚型。结果快速尿素酶反应、病理检查和PCR三者均阳性的标本52例,其中感染Hp vacA s1as1c／m2、s1as1c／m1T亚型菌株的阳性率分别是82．7％（43／52）、9．6％（5／52）,m区不能分型占7．7％（4／52）,Hp vacA s1as1c／m2与其他基因亚型相比较,差异有统计学意义。Hp vacA s1a和s1c亚型总是同时检出,未发现Hp vacA s1b、s2、m1亚型。HpcagA^＋菌株检出率是90．4％（47／52）,cagA^-菌株检出率9、6％（5／52）,二者比较,差异有统计学意义。Hp iceA1亚型菌株单独检出率78．8％（41／52）,Hp iceA2亚型菌株单独检出率是1．9％（1／52）,Hpi ceA1和ieeA2亚型均阳性是3．8％（2／52）,iceA1和iceA2亚型均阴性的比率是15．4％（8／52）,Hp iceA1亚型阳性率与其他基因亚型相比较,差异有统计学意义。结论Hp的Hp vacA s1as1c／m^2、cagA^＋、iceA1亚型是广州地区患儿感染的Hp的优势基因亚型,Hp vacA s1as1c／m^2、cagA。和iceA1基因组合是广州地区患儿感染的Hp的优势基因亚型组合。     

幽门螺杆菌细胞毒素相关蛋白、空泡毒素基因与儿童胃十二指肠疾病的关系

目的　探讨幽门螺杆菌 (Hp)细胞毒素相关蛋白 (cagA)、空泡毒素 (vacA)与儿童胃十二指肠疾病类型及胃黏膜炎症程度的关系。方法　采用免疫印迹法测定 88例Hp相关性慢性胃炎 (CG)、4 6例Hp相关性消化性溃疡 (PU)患儿血清cagA、vacA抗体 ,并观察胃黏膜病理变化。 结果　 1.CagA、vacA抗体总检出率分别为 85 .0 7%、91.0 4 % ,cagA抗体在CG与PU中检出率分别为 84 .0 9%与 86 .96 % ,两组相比无显著性差异 (P>0 .0 5 ) ;vacA抗体在CG与PU中检出率分别为 89.77%与 93.4 8% ,两组相比无显著性差异 (P >0 .0 5 )。 2 .134例Hp感染患儿cagA、vacA抗体均阳性 (Ⅰ型菌 ) 113例 ,cagA、vacA抗体均阴性 (Ⅱ型菌 ) 11例 ,cagA、vacA抗体仅一项阳性 (中间型 ) 10例 ,各型菌株在CG、PU中的检出率无明显差异 (P均 >0 .0 5 )。 3.113例Hp Ⅰ型菌株感染引起胃黏膜中重度炎症占 88.5 0 % ,而Ⅱ型菌株、中间型菌株感染引起胃黏膜中重度炎症分别为 4 5 .4 5 %及 5 0 .0 0 % ,两组有显著性差异 (P <0 .0 1)。结论　CagA、vacA与CG、PU发病均有关 ,不能作为区分Hp感染致不同胃十二指肠疾病的特异性指标。本地区儿童感染的Hp主要为cagA和vacA阳性的Ⅰ型菌株 ,能引起胃黏膜较重的炎症     

具有cagA、vacA基因的幽门螺杆菌感染及其与胃十二指肠疾病的关系

目的 研究上海地区儿童慢性胃炎及消化性溃疡患者中具有cagA、vacA基因幽门螺杆菌感染（HP）的感染状况以及cagA、cacA基因的存在与不同种类胃十二指肠疾病发生的关系。方法 对124例有消化道症状，年龄在4－13岁的儿童行胃镜检查，并在胃窦部取活检粘膜作HP的分离培养。利用聚合酶链反应技术（PCR）测定分离培养出的HP菌株的cagA、vacA基因进行分型。扩增所用引物：cagA1：5′－CCGGAGAATTCGATAACAGGCAAGCTTTTGAGG－3′，cag2：5′－GCCTGCAGTTATCGAAAA－GATTGTTTGGCAG－3′，vacA1：5′－GTCAGCATCACACCGCAAC－3′，vacA2：5′－CTGCTTGASATGCGCCAAAC－3′。结果 124例患儿中，分离培养出的HP菌株61株，平均检出率为50％，其中慢性胃炎培养阳性率为48．45％（47／97例），十二指肠球部溃疡为62．30％（14／26例），基因测定结果显示，61株HP中62．30％含有cagA基因，42．62％含有vascA基因。慢性胃炎和二十指肠球部溃疡cagA基因检出率相似（分别为61．70％和62．28％），而vacA基因检出率十二指肠球部溃疡明显高于慢性胃炎组（71．43％和34．04％，χ^2＝6．166，P＜0．05）。进一步分型发现感染Hp菌株的十二指肠球部胃炎组（71．43％和34．04％，χ^2＝6．166，P＜0．05）。进一步分型发现Hp菌株的十二肠球部溃疡患儿中，50％（7／14）为cagA^ 、vacA^ 型，慢性胃炎患儿中330％（18／47）为cagA^ 、vacA^-型Hp菌株，统计学检查差异有显著意义（χ^2＝13．48，P＜0．05），提示vacA与十二指肠球部溃疡的发生密切相关。结论 十二指肠球部溃疡患儿感染的HP多为cagA^ 、vacA^ 的I型菌，vacA是致小儿十二指肠溃疡的重要因素，vagA与慢性胃炎、十二指肠溃疡的发生有关，但不能作为区分HP感染致不同胃肠道疾病的单一指标。     

幽门螺杆菌细胞毒素相关基因、空泡毒素基因及其表达产物与儿童胃十二指肠疾病的关系

目的　研究幽门螺杆菌 (Hp)的细胞毒素相关基因 (cagA基因 )、空泡毒素基因 (vacA基因 )及其表达产物cagA蛋白、vacA抗体与儿童消化性溃疡 (PU)、慢性胃炎 (CG)之间的关系。 方法　对 15 2例Hp感染患儿 (PU 31例 ,CG 12 1例 )的胃黏膜应用 4对不同引物 ,通过聚合酶链反应 (PCR)技术检测cagA、vacA基因 ;用免疫印迹法测定 139例血清cagA、vacA抗体。结果　 15 2例胃黏膜 10 0 %具有vacA基因 ( 15 2 /15 2 ) ,cagA基因经用 3对不同引物检测 ,其总阳性率为 78% ( 118/15 2 ) ,其中PU 71% ( 2 2 /31) ,CG 79% ( 96 /12 1) ,两组间差异无显著性 ( χ2 =0 .996　P >0 .0 5 )。 139例血清vacA抗体阳性率 92 .1% ( 12 8/139) ,PU 86 .7% ( 2 6 /30 ) ,CG 94 .5% ( 10 3/10 9) ( χ2 =1.14 6　P >0 .5 ) ;cagA抗体阳性率 91.4 % ( 12 7/139) ,PU 83.3% ( 2 5 /30 ) ,CG93.6 % ( 10 2 /10 9) ( χ2 =3.130　P >0 .5 ) ,以上两组间差异均无显著性。结论　 1.浙江地区儿童感染的Hp绝大多数为具有cagA基因的毒力菌株。 2 .亚洲地区与西方人群中感染的Hp菌株可能存在等位基因的多态性、变异性     

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目的探讨儿童幽门螺杆菌vacA基因型并分析各基因亚型与胃十二指肠疾病的关系。方法对80例H·pylori感染的儿童,采用PCR法扩增胃黏膜vacA基因亚型。结果南宁儿童H·pylorivacA基因型有s1a/m1和s1a/m2两种组合,基因频率分别为3·75%、66·25%。vacA基因各亚型在慢性浅表性胃炎及消化性溃疡中的检出率差异无显著性(P>0·05)。结论s1a/m2为南宁地区儿童幽门螺杆菌的vacA优势基因型。部分患儿同时感染多株不同vacA基因型H·pylori。vacA基因各亚型不能作为南宁地区儿童H·pylori菌株毒力强弱的指标。     

幽门螺杆菌菌株类型与儿童上消化道疾病的关系

目的：幽门螺杆菌（Helicobacter pylori,Hp）感染已被确立为是引起慢性浅表性胃炎和消化性溃疡的重要病因，是胃癌和胃黏膜相关性淋巴样组织（MALT）淋巴瘤的重要危险因素。Hp的致病性与毒力有关，而细胞毒素相关蛋白（CagA）和空泡毒素（VcaA）是Hp的主要毒力因子之一。该研究通过了解Hp菌株类型与儿童胃十二指肠疾病类型及胃窦黏膜病理组织学变化的关系，探讨Hp感染的分型诊断是否有助于判断儿童胃十二指肠疾病的严重程度。方法：采用免疫印迹法对115例有上消化道症状的患儿进行Hp的血清学分型，并行胃镜检查，观察胃十二指肠疾病类型。取胃窦黏膜经Harris配方苏木精染色观察胃窦黏膜病理组织学变化、亚甲基蓝染色观察Hp感染情况。结果：115例患儿中检出Hp Ⅰ型菌株84例（73.0%），中间型菌株21例（18.3%），Ⅱ型菌株10例（8.7%）；Ⅰ型菌株引起胃窦黏膜中、重度炎症分别为83例、1例；中间型菌株引起胃窦黏膜中度炎症21例；Ⅱ型菌株引起胃窦黏膜轻度炎症2例，中度炎症8例，经统计学处理，各型菌株在引起胃窦黏膜炎症程度上差异有显著性（χ2=15.444，P<0.01），Ⅰ型菌株感染引起胃窦黏膜炎症程度最重，Ⅱ型菌株感染引起胃窦黏膜炎症程度最轻；而在活动性、萎缩的发生率上，各型差异无显著性（P＞0.05）；在淋巴滤泡形成的发生率上，各型差异有显著性（χ2=10.171，P<0.01）。各型菌株引起胃镜下胃、十二指肠疾病类型的构成比无明显差异（P＞0.05）。结论：该地区儿童Hp感染以Ⅰ型菌株最为多见。各型菌株100%存在胃窦黏膜组织学改变。Ⅰ型菌株感染所致胃窦黏膜炎症程度最重，且引起淋巴滤泡形成的发生率最高。Hp感染的分型诊断无助于对儿童胃、十二指肠疾病类型的判断，但有助于对儿童胃、十二指肠疾病病情的判断，Ⅰ型菌株感染者需要更为积极的治疗，对于Hp感染的儿童无论其血清分型如何，均应引起重视，并长期随访。［中国当代儿科杂志，2007，9（3）：201-204］     

儿童幽门螺杆菌细胞毒素相关基因A检测的意义

目的探讨儿童幽门螺杆菌(Hp)细胞毒素相关基因A(cagA)的流行情况,分析其与胃十二指肠疾病的关系。方法对80例患儿经胃黏膜快速尿素酶试验和Warthin-Starry银染色,聚合酶链式反应(PCR)法检测Hp尿素酶C基因及血清抗Hp-IgG确诊为Hp感染儿童采用PCR法检测胃黏膜cagA。结果80例患儿胃黏膜Hp菌株应用cagA的5对不同引物分别扩增ca-gA DNA 400、280、349、298 bp和191 bp 5个片段,cagA检出率分别为43.8%、20.0%、55.0%、32.5%和61.2%,它们之间有显著性差异。cagA总检出率为83.7%。轻度与中重度炎症cagA检出率无显著性差异。慢性浅表性胃炎及消化性溃疡两组cagA检出率无显著性差异。结论南宁地区儿童Hp以cagA阳性为主。cagA基因与胃黏膜炎症程度无关,且cagA基因不能作为南宁地区Hp菌株毒力强弱的指标。     

胃十二指肠疾病儿童幽门螺杆菌cagA、vacA和iceA基因型分布

目的了解来自于江西地区胃十二指肠疾病儿童感染的幽门螺杆菌(H.pylori)临床分离菌株cag A、vac A和ice A基因亚型分布情况,并探讨H.pylori cag A、vac A和ice A基因亚型与儿童胃十二指肠疾病类型之间的关系。方法从来自江西地区的316例患有胃十二指肠疾病儿童的胃窦黏膜中,培养出107株H.pylori菌株,提取菌株基因组DNA,采用PCR法检测H.pylori ure A、cag A、vac A及ice A基因亚型。结果在107株H.pylori临床分离菌株中,H.pylori ure A基因和cag A基因检出率分别为100%(107/107)和94.4%(101/107)。vac A基因总检出率为100%(107/107),vac As1a、vac As1c、vac Am1和vac Am2基因检出率分别为74.8%(80/107)、25.2%(27/107)、29.9%(32/107)和69.2%(74/107),其中0.9%(1/107)H.pylori菌株同时检测出vac Am1和vac Am2基因型;在vac A基因的嵌合体中,vac As1a/m1、vac As1a/m2、vac As1c/m1和vac As1c/m2基因检出率分别为26.2%(28/107)、51.4%(55/107)、3.7%(4/107)和17.8%(19/107)(P0.001)。ice A1和ice A2基因阳性检出率分别为79.4%(85/107)和9.3%(10/107)(P0.001);ice A1和ice A2基因亚型均阳性的检出率为7.5%(8/107)。H.pylori各基因亚型在消化性溃疡、慢性胃炎和十二指肠球炎3组间的检出率比较差异无统计学意义(P0.05)。结论来自于江西地区胃十二指肠疾病儿童感染的H.pylori优势基因亚型是cag A、vac As1a/m2和ice A1;H.pylori感染存在不同基因型菌株混合感染;H.pylori基因亚型与胃十二指肠疾病类型无相关性。     

不同菌株类型幽门螺杆菌感染胃肠黏膜的早期病理组织学改变

目的通过对不同菌株类型幽门螺杆菌(Hp)感染儿童胃肠黏膜病理组织学改变进行对比研究,了解其早期致病性。方法采用Western Blot免疫印迹法对Hp阳性72例及Hp阴性36例患儿进行Hp血清学菌株分型,并参照悉尼胃炎分类标准,对Hp菌株类型感染患儿胃肠黏膜进行病理组织学对比研究。通过黏膜病理炎症程度、活动性、淋巴滤泡形成及上皮化生等方面进行评价。结果本组HpⅠ型高毒力菌株检出率68.1%(49例),中间型Hp菌株27.7%(20例),Ⅱ型低毒力菌株4.2%(3例)。对Hp毒力菌株进行病理组织学观察,发现HpⅠ型与中间型菌株在致胃窦黏膜组织炎症活动性方面,有显著性差异(P<0.01);HpⅠ型较中间型菌株在致十二指肠球部黏膜组织炎性及炎症活动性方面均明显增强(P均<0.01)。HpⅡ型感染患儿3例,1例为窦部、2例为球部轻度炎症,均无活动性炎性表现。HpⅠ型高毒力菌株至胃窦黏膜淋巴滤泡发生率44.8%,较中间型(15.0%)显著增高(P<0.05)。HpⅠ型与中间型菌株感染黏膜萎缩、肠上皮化生方面无显著差异。结论本地区儿童Hp感染菌株类型以Ⅰ型高毒力菌株为主,同时存在中间型及Ⅱ型低毒力菌株感染。Hp感染早期阶段已出现胃肠黏膜活动性炎性改变。     

幽门螺杆菌毒素因子与儿童胃十二指肠疾病的相关性研究

目的　研究贵阳地区儿童幽门螺杆菌感染的CagA蛋白及VacA蛋白的感染状况 ,以及不同Hp菌株类型与胃肠黏膜病理组织学改变的关系。方法　采用免疫印迹法 (WesternBlot)对 6 7例Hp阳性及 32例Hp阴性患儿血清进行Hp毒素因子的测定及血清学菌株分型 ,参照悉尼胃炎分类标准 ,对不同Hp菌株类型进行病理组织学对比研究。结果　本地区Ⅰ型高毒力Hp菌株检出率 6 8.6 % ,中间型Hp菌株为 2 6 8% ,Ⅱ型Hp低毒力菌株为 4 4 %。Ⅰ型Hp菌株 1 0 0 %存在胃肠黏膜组织学改变 ,胃窦黏膜以中度炎症为主 ,占 73 9% ,重度炎症占 1 1 % ,活动性占 73 9%。十二指肠球部黏膜以中 重度炎症为主 ,占77 3% ,活动性占 6 6 4 %。中间型Hp菌株 72 %存在黏膜组织学改变 ,胃窦黏膜以轻度炎症为主 ,占 5 5 6 % ,活动性占 5 5 % ,十二指肠球部黏膜以轻 中度炎症为主 ,占 5 5 1 % ,活动性占 1 1 1 %。Ⅰ型Hp菌株与中间型Hp菌株在致黏膜炎症改变程度及活动性方面有差异性 (P <0 0 5 )。在淋巴滤泡形成方面 ,Ⅰ型Hp菌株较中间型Hp菌株有显著性差异 (P <0 0 1 )。结论　本地区Hp感染菌株类型以Ⅰ型为主 ,占 6 8 6 % ,同时存在 2 6 8%的中间型菌株及 4 4 %的Ⅱ型菌株的感染。Ⅰ型Hp菌株及中间型菌株在黏膜炎症程度及活动性上有差异性     

Helicobacter pylori genotypes in Israeli children: the significance of geography

BACKGROUND: There is substantial genetic variation among different isolates of Helicobacter pylori, which may affect the clinical outcome. The aims of this study were to find the common H. pylori genotypes in Israeli children and to look for a possible genotype-phenotype correlation. METHODS: Ninety-eight H. pylori cultures were isolated from antral biopsy specimens of symptomatic Israeli children and were analyzed for vacA and iceA genotype and cagA and cagE status by polymerase chain reaction. RESULTS: cagA and cagE genes were present in only 25.5% and 24.5%, respectively. The common vacA genotype was s2m2, which was found in 65%. Eleven specimens (11%) contained multiple vacA genotypes. iceA1 was found in 37% and iceA2 in 52% of cases. Both iceA alleles were found in 11%. Increased prevalence of iceA1 and cagE were observed in children with duodenal disease, although it did not reach significance. CONCLUSIONS: The low prevalence of cagA and the high prevalence of vacA genotype s2m2 in Israeli pediatric patients are different from the genotype prevalence reported globally. However, similar findings have been reported in Egypt, indicating a possible geographic influence. There is a possible correlation between duodenal ulcer and cag E and ice A1 genotype, but the power of the study was too low to prove it.     

cagA,vacA, and iceA genotypes of Helicobacter pylori in Korean children

Background: Several putative virulence factors for Helicobacter pylori have been identified including cagA, vacA, and iceA. The aims of the present study were to study the distribution of cagA, vacA, and iceA genotypes in children with H. pylori gastritis and to examine the association of genotypes with severity of gastritis. Methods: H. pylori DNA was extracted from antral biopsy specimens from 33 children with H. pylori gastritis. Specific polymerase chain reaction assays were used for three genes: cagA, vacA, and iceA. The features of gastritis were graded in accordance with the updated Sydney System. Results: Of the 33 children, 31 (94%) were cagA positive. Twenty‐four (72%) had s1c genotype and nine (27%) had s1a. The m1 genotype was seen in 27 (82%) and m2 was found in five (15%). The iceA1 genotype was detected in 25 (76%). Scores of neutrophil activity, chronic inflammation, and H. pylori density were independent of cagA, vacA and iceA status. Conclusion: The cagA‐positive vacA s1c/m1 iceA1 genotype was predominant in Korean children with recurrent abdominal pain and H. pylori gastritis. The cagA, vacA and iceA genotype were not associated with the severity of gastritis.     

Helicobacter pylori infection, histopathological gastritis and gastric epithelial cell apoptosis in children

AIM: Features of gastritis and gastric epithelial cell apoptosis in children infected with Helicobacter pylori genotypes are seldom studied. Therefore, we investigated the relationship between vacA genotypes and the severity of gastritis, and gastric epithelial cell apoptosis in H. pylori-infected children. METHODS: Antral biopsies from 52 children infected with H. pylorivacA genotypes (s1a/m1 = 17, s1a/m2 = 21 and s2/m2 = 14) were analysed for severity of gastritis on histopathology. Fifteen biopsies infected with different vacA genotypes were studied for gastric epithelial cell apoptosis by terminal uridine deoxynucleotidyl nick-end labelling. RESULTS: Children infected with the s1a/m1 and s1a/m2 vacA genotypes had higher severity of chronic inflammation than the s2/m2 genotype (s1a/m1 vs s2/m2, p=0.05; s1a/m2 vs s2/m2, p=0.01). The vacA s1a allele was more independently associated with severe chronic inflammation than the s2 allele (p=0.02). Children infected with the s1a/m1 and s1a/m2 strains had higher gastric epithelial cell apoptosis than the s2/m2 strain (s1a/m1 or s1a/m2 vs s2/m2, p<0.0001). CONCLUSION: The s1a/m1 and s1a/m2 H. pylorivacA genotypes have significantly higher association with severe chronic gastritis and gastric epithelial cell apoptosis than the s2/m2 genotype in children. The role of H. pylorivacA genotypes and their allelic subtypes in relation to pathogenicity and disease potential in children needs further studies.     

幽门螺杆菌感染儿童厌食机制研究

目的探讨Ghrelin表达以及不同幽门螺杆菌(H.pylori)毒力基因型在H.pylori感染患儿厌食发病机制中的作用。方法 H.pylori感染患儿60例,根据临床表现分为厌食(n=30例)和非厌食(n=30例)组。应用RT-PCR方法检测胃黏膜Ghrelin mRNA表达水平,比较两组患儿以及厌食患儿在H.pylori根治前后的差异。同时采用PCR方法检测所有患儿的H.pylori毒力cagA/vacA基因并分型。结果 H.pylori感染厌食患儿胃黏膜GhrelinmRNA表达低于非厌食患儿,两组差异有统计学意义(P<0.01);厌食患儿H.pylori根治后Ghrelin mRNA表达明显升高,与根治前比较差异有统计学意义(P<0.05);同时患儿食欲改善,体质量增长显著。厌食与非厌食患儿感染的H.pylori均为Ⅰ型毒力株,厌食患儿的cagA m1阳性率高于非厌食患儿,厌食患儿的H.pylori基因型以s1/m1多见。结论 Ghrelin在H.pylori感染厌食患儿中表达降低,而H.pylori根治后表达升高,H.pylori可能通过影响Ghrelin分泌而导致厌食;毒力较强的H.pylori ...     

IL1RN polymorphism and cagA-positive Helicobacter pylori strains increase the risk of duodenal ulcer in children

Duodenal ulcers in children are associated with Helicobacter pylori gastric infection with cagA-positive strains, but factors linked to the host are poorly known. The authors evaluated the role of proinflammatory interleukin-1 gene cluster polymorphisms in the pathogenesis of duodenal ulcer. They studied prospectively 437 children 1 to years old, 209 of whom were H. pylori positive and 228 of whom were H. pylori negative. IL1B-511-C/T, -31T/C, and IL1RN Variable number of tandem repeats were genotyped by polymerase chain reaction (PCR) restriction fragment length polymorphism, PCR with confronting two-pair primers, and PCR, respectively. cagA status was evaluated by PCR. The role of the proinflammatory cytokine genotypes in the genesis of duodenal ulcer was evaluated before and after stratification of H. pylori status on logistic regression models. In the group of children without duodenal ulcer, no association was observed between H. pylori status and proinflammatory polymorphisms. Furthermore, no association between IL1 cluster genotypes and cagA status was seen in the H. pylori-positive children. However, increasing age, male sex, and IL1RN*2 were independently associated with duodenal ulcer. After stratification, in the H. pylori-positive children, increasing age, male sex, the presence of ILRN*2 allele, and cagA-positive status were independently associated with duodenal ulcer. The risk for the development of duodenal ulcer increased when a combined association of the presence of IL1RN*2 allele and infection by a cagA-positive H. pylori strain was the variable. This study provides evidence supporting independent roles of IL1RN*2 allele and cagA-positive status in the genesis of duodenal ulcer in children.