幽门螺杆菌毒力基因与克拉霉素耐药基因突变相关性
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摘要
目的分析浙江省台州地区幽门螺杆菌(helicobacter pylori,Hp)cag A和Vac A为主的毒力基因型分布,探讨其与克拉霉素耐药基因突变类型的关系。方法招募存在上消化道症状患者,并行胃镜检查活检患者胃黏膜组织。通过Hp分离培养方法,培养出216株Hp。采用聚合酶链式反应(polymerase chain reaction,PCR)检测并分析cag A和Vac A的分型情况。采用PCR扩增和Sanger测序技术检测克拉霉素耐药基因(23S rRNA)2142和2143位碱基点突变情况。结果 Cag A基因的阳性检出率为99.54%(215/216),其中Cag A-D为95.83%(207/216)、Cag A-C为3.70%(8/216)。Vac A基因阳性检出率为97.22%(210/216),s1、s2、m1、m2亚型分别为97.22%(210/216)、0、35.19%(76/216)、60.65%(131/216)。本研究发现,浙江省台州地区毒力基因以Cag A-D和Vac As1m2型为优势基因型。此外,23S rRNA基因2142和2143位点均未突变的有93株。突变的有123株,其中A2142G位点突变4株、A2143G位点突变119株。毒力基因型与23S rRNA基因2142和2143位点突变类型无相关性(P>0.05)。结论台州地区为HP高毒力地区,且以Cag A-D和Vac As1m2型为主,其分布与克拉霉素耐药基因突变无密切相关性。
Objective To analyze the distributions of virulence genes cag A and Vac A from helicobacter pylori( Hp) and to study the relationship between virulence genotype and gene mutation associated with clarithromycin resistance. Methods A total of 216 Hp were isolated from gastric mucosal tissue in patients with upper gastrointestinal symptoms. The cag A and Vac A genotypes were detected and analyzed by polymerase chain reaction( PCR). The mutations at positions 2142 and 2143 of clarithromycin resistance gene( 23 S rRNA)were detected by PCR and Sanger sequencing. Results The positive rate of Cag A gene was 99. 54%( 215/216),of which Cag A-D was95. 83%( 207/216) and Cag A-C was 3. 70%( 8/216). The positive rate of Vac A gene was 97. 22%( 210/216),and the s1,s2,m1,and m2 subtypes were 97. 22%( 210/216),0,35. 19%( 76/216) and 60. 65%( 131/216),respectively. In this study,Cag A-D and Vac As1 m2 were dominant genotypes in Taizhou region. Additionally,there were 93 Hp strians which were not mutated in 2142 and2143 gene loci of 23 S rRNA. There were 123 Hp mutants,of which 4 strains were A2142 G mutations and 119 strains were A2143 G mutations. There were no relationship between virulence genotype and 2142 and 2143 gene loci of 23 S rRNA mutation types( P > 0. 05). Conclusion Taizhou is a region with high Hp toxicity and is mainly in Cag A-D and Vac As1 m2. The distribution of the Hp toxicity is not closely related to the clarithromycin-resistance gene mutation.
Objective To analyze the distributions of virulence genes cag A and Vac A from helicobacter pylori( Hp) and to study the relationship between virulence genotype and gene mutation associated with clarithromycin resistance. Methods A total of 216 Hp were isolated from gastric mucosal tissue in patients with upper gastrointestinal symptoms. The cag A and Vac A genotypes were detected and analyzed by polymerase chain reaction( PCR). The mutations at positions 2142 and 2143 of clarithromycin resistance gene( 23 S rRNA)were detected by PCR and Sanger sequencing. Results The positive rate of Cag A gene was 99. 54%( 215/216),of which Cag A-D was95. 83%( 207/216) and Cag A-C was 3. 70%( 8/216). The positive rate of Vac A gene was 97. 22%( 210/216),and the s1,s2,m1,and m2 subtypes were 97. 22%( 210/216),0,35. 19%( 76/216) and 60. 65%( 131/216),respectively. In this study,Cag A-D and Vac As1 m2 were dominant genotypes in Taizhou region. Additionally,there were 93 Hp strians which were not mutated in 2142 and2143 gene loci of 23 S rRNA. There were 123 Hp mutants,of which 4 strains were A2142 G mutations and 119 strains were A2143 G mutations. There were no relationship between virulence genotype and 2142 and 2143 gene loci of 23 S rRNA mutation types( P > 0. 05). Conclusion Taizhou is a region with high Hp toxicity and is mainly in Cag A-D and Vac As1 m2. The distribution of the Hp toxicity is not closely related to the clarithromycin-resistance gene mutation.
引文
1 Diaconu S,Predescu A,Moldoveanu A,et al.Helicobacter pylori infection:old and new[J].J Med Life,2017,10(2):112-117
2 Herrera AG.Helicobacter pylori and food products:a public health problem[J].Methods Mol Biol,2004,268:297-301
3 刘文忠,谢勇,陆红等.第五次全国幽门螺杆菌感染处理共识报告[J].胃肠病学,2017,22(6):346-360
4 BAA Talebi,MA Mohabbati.High prevalence of helicobacter pylori hop Q II genotype isolated from Iranian patients with gastroduodenal disorders[J].J Pathogens,2016,2014(2014):842469
5 Kim SY,Choi DJ,Chung JW.Antibiotic treatment for Helicobacter pylori:is the end coming?[J].World J Gastrointest Pharmacol Ther,2015,6(4):183-198
6 Malfertheiner P,Megraud F,O'morain CA,et al.Management of helicobacter pylori infection——the Maastricht IV/florence consensus report[J].Gut,2012,61(5):646-664
7 杨宁敏,孟飞,徐书华,等.基于浙江省幽门螺杆菌耐药监测大数据下的根除治疗对策研究[J].中华消化内镜杂志,2016,33(11):738-742
8 Eghbali Z,Mojtahedi A,Moien A M,et al.Detection of 23SrRNA mutations strongly related to clarithromycin resistance in helicobacter pylori strains isolated from patients in the North of Iran[J].Jundishapur J Microbiol,2016,9(2):e29694
9 Suzuki RB,Lopes RA,Da CLG,et al.Low Helicobacter pylori primary resistance to clarithromycin in gastric biopsy specimens from dyspeptic patients of a city in the interior of Sao Paulo,Brazil[J].BMC Gastroenterology,2013,13(1):164
10 Boyanova L,Markovska R,Yordanov D,et al.Clarithromycin resistance mutations in helicobacter pylori in association with virulence factors and antibiotic susceptibility of the strains[J].Microb Drug Resist,2016,22(3):227-232
11 林定赛,夏莲,方蕾,等.幽门螺杆菌Vac A基因型与耐药性相关性研究[J].医学研究杂志,2018,47(2):124-126
12 刘丽娜,陈庆明.不同消化性疾病患者幽门螺杆菌毒力基因类型及意义研究[J]国际检验医学杂志,2017,38(15):2085-2087
13 范聪聪,王丽波,江海洋,等.长春地区幽门螺杆菌cag A基因及EPIYA基序多态性分析[J].中华微生物学和免疫学杂志,2017,37(6):431-437
14 Shiota S,Suzuki R,Yamaoka Y.The significance of virulence factors in Helicobacter pylori[J].J Dig Dis,2013,14(7):341-349
15 江海洋,王丽波,范聪聪,等.幽门螺杆菌毒力基因cag A、vac A和ice A与胃十二指肠疾病的关系[J].中国微生态学杂志,2017,29(3):256-260
16 Li Q,Liu J,Gong Y,et al.Association of Cag A EPIYA-D or EPIYA-Cphosphorylation sites with pepticμlcer and gastric cancer risks:a meta-analysis[J].Medicine(Baltimore),2017,96(17):e6620
17 Fahimi F,Tohidkia MR,Fouladi M,et al.Pleiotropic cytotoxicity of Vac A toxin in host cells and its impact on immunotherapy[J].Bioimpacts Bi,2017,7(1):59-71
18 林朗,杨恩,张红河,等.浙江省幽门螺杆菌cag A、vac A基因型的流行病学研究[J].医学研究杂志,2008,39(6):94-96
19 Kyrillos A,Arora G,Murray B,et al.The presence of phage orthologous genes in helicobacter pylori correlates with the presence of the virulence factors Cag A and Vac A[J].Helicobacter,2016,21(3):226-233
20 Garcia GT,Aranda KR,Gon9alves ME,et al.High prevalence of clarithromycin resistance and cag A,vac A,ice A2,andbab A2 genotypes of Helicobacter pylori in Brazilian children[J].J Clin Microbiol,2010,48(11):4266-4268
2 Herrera AG.Helicobacter pylori and food products:a public health problem[J].Methods Mol Biol,2004,268:297-301
3 刘文忠,谢勇,陆红等.第五次全国幽门螺杆菌感染处理共识报告[J].胃肠病学,2017,22(6):346-360
4 BAA Talebi,MA Mohabbati.High prevalence of helicobacter pylori hop Q II genotype isolated from Iranian patients with gastroduodenal disorders[J].J Pathogens,2016,2014(2014):842469
5 Kim SY,Choi DJ,Chung JW.Antibiotic treatment for Helicobacter pylori:is the end coming?[J].World J Gastrointest Pharmacol Ther,2015,6(4):183-198
6 Malfertheiner P,Megraud F,O'morain CA,et al.Management of helicobacter pylori infection——the Maastricht IV/florence consensus report[J].Gut,2012,61(5):646-664
7 杨宁敏,孟飞,徐书华,等.基于浙江省幽门螺杆菌耐药监测大数据下的根除治疗对策研究[J].中华消化内镜杂志,2016,33(11):738-742
8 Eghbali Z,Mojtahedi A,Moien A M,et al.Detection of 23SrRNA mutations strongly related to clarithromycin resistance in helicobacter pylori strains isolated from patients in the North of Iran[J].Jundishapur J Microbiol,2016,9(2):e29694
9 Suzuki RB,Lopes RA,Da CLG,et al.Low Helicobacter pylori primary resistance to clarithromycin in gastric biopsy specimens from dyspeptic patients of a city in the interior of Sao Paulo,Brazil[J].BMC Gastroenterology,2013,13(1):164
10 Boyanova L,Markovska R,Yordanov D,et al.Clarithromycin resistance mutations in helicobacter pylori in association with virulence factors and antibiotic susceptibility of the strains[J].Microb Drug Resist,2016,22(3):227-232
11 林定赛,夏莲,方蕾,等.幽门螺杆菌Vac A基因型与耐药性相关性研究[J].医学研究杂志,2018,47(2):124-126
12 刘丽娜,陈庆明.不同消化性疾病患者幽门螺杆菌毒力基因类型及意义研究[J]国际检验医学杂志,2017,38(15):2085-2087
13 范聪聪,王丽波,江海洋,等.长春地区幽门螺杆菌cag A基因及EPIYA基序多态性分析[J].中华微生物学和免疫学杂志,2017,37(6):431-437
14 Shiota S,Suzuki R,Yamaoka Y.The significance of virulence factors in Helicobacter pylori[J].J Dig Dis,2013,14(7):341-349
15 江海洋,王丽波,范聪聪,等.幽门螺杆菌毒力基因cag A、vac A和ice A与胃十二指肠疾病的关系[J].中国微生态学杂志,2017,29(3):256-260
16 Li Q,Liu J,Gong Y,et al.Association of Cag A EPIYA-D or EPIYA-Cphosphorylation sites with pepticμlcer and gastric cancer risks:a meta-analysis[J].Medicine(Baltimore),2017,96(17):e6620
17 Fahimi F,Tohidkia MR,Fouladi M,et al.Pleiotropic cytotoxicity of Vac A toxin in host cells and its impact on immunotherapy[J].Bioimpacts Bi,2017,7(1):59-71
18 林朗,杨恩,张红河,等.浙江省幽门螺杆菌cag A、vac A基因型的流行病学研究[J].医学研究杂志,2008,39(6):94-96
19 Kyrillos A,Arora G,Murray B,et al.The presence of phage orthologous genes in helicobacter pylori correlates with the presence of the virulence factors Cag A and Vac A[J].Helicobacter,2016,21(3):226-233
20 Garcia GT,Aranda KR,Gon9alves ME,et al.High prevalence of clarithromycin resistance and cag A,vac A,ice A2,andbab A2 genotypes of Helicobacter pylori in Brazilian children[J].J Clin Microbiol,2010,48(11):4266-4268